U.S. patent application number 13/037575 was filed with the patent office on 2011-06-23 for novel aromatic compounds and their use in medical applications.
This patent application is currently assigned to REVOTAR BIOPHARMACEUTICALS AG. Invention is credited to Ewald M. AYDT, Anke S. Busemann, Remo Kranich.
Application Number | 20110152291 13/037575 |
Document ID | / |
Family ID | 35717680 |
Filed Date | 2011-06-23 |
United States Patent
Application |
20110152291 |
Kind Code |
A1 |
AYDT; Ewald M. ; et
al. |
June 23, 2011 |
NOVEL AROMATIC COMPOUNDS AND THEIR USE IN MEDICAL APPLICATIONS
Abstract
Pharmaceutical compositions comprising at least one compound of
the formulas (Ia) or (Ib) and a pharmaceutically acceptable carrier
wherein the symbols have the following meaning --X- is e.g. and Y
being e.g. or the pharmaceutically acceptable salts can be applied
to modulate the in-vitro and in-vivo binding processes mediated by
E-, P- or L-selectin binding.
Inventors: |
AYDT; Ewald M.; (Berlin,
DE) ; Kranich; Remo; (Berlin, DE) ; Busemann;
Anke S.; (Henningsdorf, DE) |
Assignee: |
REVOTAR BIOPHARMACEUTICALS
AG
Hennigsdorf
DE
|
Family ID: |
35717680 |
Appl. No.: |
13/037575 |
Filed: |
March 1, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12067389 |
May 1, 2008 |
7923473 |
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PCT/EP2006/009154 |
Sep 20, 2006 |
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13037575 |
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Current U.S.
Class: |
514/255.01 ;
435/7.1; 514/256; 514/382; 514/438; 514/471; 514/539; 514/622;
544/329; 544/391; 548/252; 549/486; 549/77; 560/45; 564/174 |
Current CPC
Class: |
A61P 25/00 20180101;
A61P 19/02 20180101; A61P 9/10 20180101; A61P 37/02 20180101; A61P
11/00 20180101; A61P 37/08 20180101; A61P 11/06 20180101; A61P
29/00 20180101; A61P 43/00 20180101; A61K 31/05 20130101; A61P 9/00
20180101; A61P 31/04 20180101; A61P 17/06 20180101; A61P 1/04
20180101 |
Class at
Publication: |
514/255.01 ;
560/45; 549/77; 549/486; 544/391; 548/252; 564/174; 544/329;
514/539; 514/438; 514/471; 514/382; 514/622; 514/256; 435/7.1 |
International
Class: |
A61K 31/495 20060101
A61K031/495; C07C 235/78 20060101 C07C235/78; C07D 333/24 20060101
C07D333/24; C07D 307/54 20060101 C07D307/54; C07D 295/16 20060101
C07D295/16; C07D 409/04 20060101 C07D409/04; C07C 235/42 20060101
C07C235/42; C07D 239/42 20060101 C07D239/42; A61K 31/216 20060101
A61K031/216; A61K 31/381 20060101 A61K031/381; A61K 31/341 20060101
A61K031/341; A61K 31/41 20060101 A61K031/41; A61K 31/167 20060101
A61K031/167; A61K 31/505 20060101 A61K031/505; G01N 33/53 20060101
G01N033/53; A61P 11/00 20060101 A61P011/00; A61P 29/00 20060101
A61P029/00; A61P 17/06 20060101 A61P017/06; A61P 19/02 20060101
A61P019/02; A61P 9/00 20060101 A61P009/00; A61P 9/10 20060101
A61P009/10; A61P 11/06 20060101 A61P011/06; A61P 43/00 20060101
A61P043/00 |
Foreign Application Data
Date |
Code |
Application Number |
Sep 20, 2005 |
EP |
05020510.3 |
Claims
1. A compound of the formulas (Ia) or (Ib): ##STR00061## wherein
the symbols and substituents have the following meaning: --X-- is
##STR00062## wherein "ring" is ##STR00063## wherein R.sup.1 is H,
NO.sub.2, CF.sub.3, F, Cl, Br, I, CN, CH.sub.3, NH.sub.2, NHAlkyl,
NHAryl, NHAcyl, and k=0 or 1; ##STR00064## wherein "ring" is as
defined above; T is O, S or [H,H]; p=0, 1, or 2; --Y is
##STR00065## wherein s is 0 or 1; R.sup.2 is CO.sub.2 H,
CO.sub.2alkyl, CO.sub.2aryl, CO.sub.2NH.sub.2, CO.sub.2aralkyl,
CH.sub.2SO.sub.3 H, CH.sub.2SO.sub.2NH.sub.2,
CH.sub.2PO.sub.2(OH).sub.2, SO.sub.3 H, SO.sub.2NH.sub.2,
PO(OH).sub.2, 1-H-tetrazolyl-, CHO, COCH.sub.3, CH.sub.2OH,
CH.sub.2NH.sub.2, NH.sub.2, CH.sub.2NHalkyl,
CH.sub.2N(alkyl)alkyl', NHalkyl, N(alkyl)alkyl', OCH.sub.3,
CH.sub.2OCH.sub.3, CH.sub.2SH, SH, F, Cl, Br, I, CH.sub.3,
CH.sub.2CH.sub.3, CN, or CF.sub.3; R.sup.3 independently from
R.sup.2 is H, CH.sub.3, CH.sub.2CH.sub.3, CF.sub.3, F, Cl, Br, I,
CN, or NO.sub.2; R.sup.4 independently from R.sup.2 and R.sup.3 is
H, CH.sub.3, CH.sub.2CH.sub.3, CF.sub.3, F, Cl, Br, I, CN,
NO.sub.2, or R.sup.2; R.sup.5 is H, NO.sub.2, CF.sub.3, F, Cl, Br,
I, CN, CH.sub.3, OCH.sub.3, SH, or NH2; --W-- is
--(CH.sub.2--).sub.v, cis-CH.dbd.CH--or trans-CH.dbd.CH--, and v is
0, 1, or 2; in case that --W-- is cis-CH.dbd.CH--or
trans-CH.dbd.CH--, R.sup.2 must not be NH.sub.2 or SH; ##STR00066##
wherein t is 0, 1, or 2; --Z is ##STR00067## R.sup.7 independently
from R.sup.2 is H, NO.sub.2, CF.sub.3, F, Cl, Br, I, CN, CH.sub.3,
OCH.sub.3, SH, or NH.sub.2; ##STR00068## in which K is NH, NMe, O,
or S; or a pharmaceutically acceptable salt, ester, or amide
thereof of the above identified compounds of formula (Ia) or
(Ib).
2. The compound of claim 1 defined by formula (IIa) or (IIb)
##STR00069## wherein --Y is as defined in claim 1 and --X'-- is
X(c) as defined in claim 1, wherein all indices, symbols, and
substituents are as defined in claim 1.
3. The compound of claim 1 defined by formulas (A1), (B1), (A2), or
(B2): ##STR00070## wherein --X'-- is X(c) as defined in claim 1 and
--Y is as defined in claim 1 and wherein --X'' is ##STR00071## and
wherein --Y' is ##STR00072## wherein all indices, symbols, and
substituents are as defined in claim 1.
4. The compound of claim 1 defined by formulas (C) or (D):
##STR00073## wherein --X'' is ##STR00074## and wherein --Y' is
##STR00075## wherein all indices, symbols, and substituents are as
defined in claim 1.
5. The compound of claim 1 defined by formulas (E) or (F):
##STR00076## wherein --X'' is ##STR00077## and --Y'' is
##STR00078## wherein R.sup.9 is CO.sub.2 H, CO.sub.2alkyl,
CO.sub.2aryl, CO.sub.2NH.sub.2, CO.sub.2aralkyl, CH.sub.2SO.sub.3H,
CH.sub.2SO.sub.2NH.sub.2, CH.sub.2PO.sub.2(OH).sub.2,
1-H-tetrazolyl-, CHO, COCH.sub.3, CH.sub.2OH, CH.sub.2NH.sub.2,
CH.sub.2NHalkyl, CH.sub.2N(alkyl)alkyl', CH.sub.2OCH.sub.3, or
CH.sub.2SH, wherein all indices, symbols, and substituents are as
defined in claim 1.
6. A method of treating or preventing Chronic Obstructive Pulmonary
Disease (COPD), acute lung injury (ALI), cardiopulmonary bypass,
acute respiratory distress syndrome (ARDS), Crohn's disease, septic
shock, sepsis, chronic inflammatory diseases such as psoriasis,
atopic dermatitis, and rheumatoid arthritis, and reperfusion injury
that occurs following heart attacks, strokes, atherosclerosis, and
organ transplants, traumatic shock, multi-organ failure, autoimmune
diseases like multiple sclerosis, percutaneous transluminal
angioplasty, asthma and inflammatory bowel disease, comprising the
administration of a therapeutic amount of at least one compound of
claim 1.
7. A method of treating or preventing Chronic Obstructive Pulmonary
Disease (COPD), acute lung injury (ALI), cardiopulmonary bypass,
acute respiratory distress syndrome (ARDS), Crohn's disease, septic
shock, sepsis, chronic inflammatory diseases such as psoriasis,
atopic dermatitis, and rheumatoid arthritis, and reperfusion injury
that occurs following heart attacks, strokes, atherosclerosis, and
organ transplants, traumatic shock, multi-organ failure, autoimmune
diseases like multiple sclerosis, percutaneous transluminal
angioplasty, asthma and inflammatory bowel disease, comprising the
administration of a therapeutic amount of at least one compound of
claim 4.
8. A method of treating, diagnosing, or preventing inflammatory
disorders, comprising the administration of a therapeutic amount of
at least one compound of claim 1.
9. A cosmetic or dermatological composition, comprising at least
one compound of claim 1.
10. A cosmetic composition comprising at least one compound of
claim 1 and at least one cosmetically tolerable component.
11. A dermatological composition comprising at least one compound
of claim 1 and at least one dermatologically tolerable
component.
12. A pharmaceutical composition comprising at least one compound
of claim 1 and a pharmaceutically acceptable excipient.
13. A pharmaceutical composition comprising at least one compound
of claim 2 and a pharmaceutically acceptable excipient.
14. A pharmaceutical composition comprising at least one compound
of claim 3 and a pharmaceutically acceptable excipient.
15. A pharmaceutical composition comprising at least one compound
of claim 4 and a pharmaceutically acceptable excipient.
16. A pharmaceutical composition comprising at least one compound
of claim 5 and a pharmaceutically acceptable excipient.
Description
[0001] The present invention relates to compounds, compositions and
methods for modulating the in vitro and in vivo processes mediated
by cell adhesion molecules. The disclosed small molecules comprise
trimethoxy phenyl subunits and modulate cell adhesion
molecule-mediated functions potently.
[0002] Cell-adhesion molecule-mediated functions are part of a
complex cascade leading to the migration of circulating white blood
cells (leukocytes) from the blood stream into the surrounding
tissue (transmigration). Physiologically, leukocyte transmigration
is of critical importance for homeostasis and immuno-surveillance
of living beings including humans. Lymphocytes for example, are
constitutively leaving the blood stream into lymphatic tissues in
order to patrol for harmful antigens. Under pathological
circumstances however, e.g. local or systemic inflammation and/or
injury of the vascular system, this fundamental process is
dys-regulated, at least in part, due to an increased surface
expression of E- and P-selectin. Consequently, the excessive
leukocyte transmigration leads to a pathological cellular
infiltrate with subsequent tissue damage in several clinically
relevant settings. Disease states such as Acute Lung Injury (ALI),
Acute Respiratory Distress Syndrome (ARDS), Asthma bronchiale
(asthma), Chronic Obstructive Pulmonary Disease (COPD), Psoriasis,
Rheumatoid Arthritis, and Sepsis are all associated with tissue
inflammation induced and perpetuated by pathologically activated
leukocytes infiltrating the respective tissue. In addition,
exaggerated leukocyte infiltration contributes to the pathogenesis
of Ischemic-Reperfusion Injury (IRI) associated with organ
transplantation, cardiopulmonary bypass or percutaneous
transluminal angioplasty.
[0003] To transmigrate, leukocytes must bind to the wall of the
vascular endothelium to diffuse through the cell wall of the
capillary into the surrounding tissue. Therefore, leukocytes have
to roll onto and then adhere to the endothelial cell wall (initial
rolling or "tethering"). This primary event in transmigration is
mediated by the selectin family of cell-adhesion molecules. In
addition to directly binding to the endothelium, leukocytes can
adhere to other leukocytes, leukocyte-particles, platelets or
platelet-derived particles that are already attached to the
endothelium.
[0004] The selectin family of adhesion molecules is comprised of
three structurally related calcium-dependent carbohydrate binding
cell surface proteins, E-, P- and L-selectin. E-selectin is
expressed only on inflamed endothelium, P-selectin is expressed on
inflamed endothelium as well as on platelets and L-selectin is
expressed on leukocytes. Selectins are composed of an amino
terminal lectin domain, an epidermal growth factor (EGF)-like
domain, a variable number of complement receptor-related repeats, a
hydrophobic transmembrane domain and a C-terminal cytoplasmic
domain. The binding interactions leading to the adhesion of the
leukocytes are supposed to be mediated by contact of the lectin
domain of the selectins and various carbohydrate ligands on the
surface of the leukocytes. All three selectins can bind with low
affinity to the carbohydrate sialyl Lewis.sup.x (sLe.sup.x), a
glycosyl moiety present on the surface of most leukocytes. A
structurally related glycosyl moiety, sialyl Lewis.sup.a
(sLe.sup.a), is predominantly found on the surface of cancer cells
[K. Okazaki et al., J. Surg. Res., 1998, 78(1). 78-84; R. P. McEver
et al., Glycoconjugate Journal, 1997, 14(5), 585-591]. In case of
P-selectin, a distinct high affinity glycoprotein ligand has been
described [R. P. McEver, R. D. Cummings, J. Clin. Invest., 1997,
100, 485-492], the so-called P-selectin glycoprotein ligand-1
(PSGL-1), which contributes to a high affinity selectin binding by
its sLe.sup.x moiety as well as by parts of its peptide components,
in particular sulphated tyrosine residues [R. P. McEver, Ernst
Schering Res. Found. Workshop, 2004, 44, 137-147]. PSGL-1 is one of
the most important selectin ligands binding with highest affinity
to P-selectin, but it also binds to E- and L-selectin [G.
Constantin; Drug News Perspect; 2004; 17(9); 579-586]. It is a
homodimeric sialomucin predominantly expressed on leukocytes.
[0005] In inflammatory diseases, dys-regulated transmigration is,
at least in part, mediated due to an increased cell surface
expression of E- and P-selectin. In contrast to their low basal
expression, E- and P-selectin expression is upregulated during
inflammation, leading to a substantial recruitment of leukocytes
into the inflamed tissue. Although selectin-mediated cell adhesion
is required for fighting infection, there are various situations in
which such cell adhesion is undesirable or excessive, resulting in
severe tissue damage instead of repair. In the case of many acute
as well as chronic inflammatory disorders [e.g., asthma, chronic
obstructive pulmonary disease (COPD), psoriasis, etc.], an
association between infiltration of activated leukocytes into the
tissue simultaneously with a marked elevation of tissue expression
of corresponding adhesion molecules, particularly E- and
P-selectin, has been demonstrated [Muller et al., J. Pathol., 2002,
198(2), 270-275; Di Stefano et al., Am. J. Respir. Crit. Care.
Med., 1994, 149(3) 803-810; Terajima et al., Arch. Dermatol. Res.,
1998, 290, 246-252]
[0006] Leukocyte infiltration may also play a role in inflammatory
symptoms in the course of transplant and graft rejection. Also the
process of blood clotting is further promoted by
leukocyte-leukocyte and leukocyte-platelet binding, which occurs
because leukocytes possess both L-selectin and its corresponding
ligand PSGL-1 and can thus interact with themselves via PSGL-1, and
they can also bind to platelets which carry P-selectin.
[0007] Therefore, the modulation of selectin-mediated cell adhesion
and other selectin mediated functions, e.g. leukocyte activation,
offers a promising possibility to interfere with and stop the
inflammation cascade at a very early step. Small molecule selectin
antagonists should modulate all three selectins simultaneously as
pan-selectin-antagonists to circumvent possible redundancies
between the selectins [M. Sperandio et al., Vascular Disease
Prevention, 2004, 1, 185-195].
[0008] Besides sLe.sup.x/sLe.sup.a, the natural, high affinity
ligand PSGL-1 is another template structure for the design of small
molecule selectin antagonists. As compared to sLe.sup.x/sLe.sup.a,
PSGL-1 shows high affinity for all three selectins. To find and to
detect novel small molecule drugs that compete with PSGL-1 and
PSGL-1-like ligands for selectin binding is therefore a promising
strategy to develop a novel class of effective pan-selectin
antagonists for treating inflammatory disorders. Selectin
antagonists may be designed using selectins as well as using a
ligand like PSGL-1 as a template structure, since they are intended
to modulate the binding between selectins and PSGL-1 or other
ligands with similar binding motifs.
[0009] Novel small molecule selectin antagonists could meet certain
requirements to be drug-like and to have potential oral
bioavailability. The term drug likeness is described in the
literature [Lipinski; Adv. Drug Dev. Rev., 1997, 23, 3-25]. Beside
other molecular properties, passively transported molecules are
supposed to have on average a relative molecular weight of less
than 500 in order to be drug like. According to these rules it is
common to define compounds with a relative molecular weight of less
500 or closely above that as small molecules. Compounds with
relative molecular weights above 500 are unlikely to be orally
bioavailable. Also the presence of highly polar carbohydrate
moieties or a peptidic components is not in accordance with the
concept of drug likeness [H. Ulbrich et al., Trends PharmacoL Sci.,
2003, 24(12), 640-647; D. Slee et al., J. Med. Chem., 2001, 44,
2094-2107]. The same accounts for the development of antibody-based
drugs; because they are polypeptides and so oral administration is
a problem. Moreover, the desired compounds must be stable during
the passage through the gastrointestinal tract so that they can be
ingested/absorbed latest by the cells of the small intestines. This
is not the case for most glycosidic molecules and peptidic
structures.
[0010] There have been various investigations to develop
low-molecular weight compounds with an modulatory effect on
selectin mediated processes. These compounds include disalicylates
and disalicylate-based C-glycosides [WO 99/29706], benzyl amino
sulfonic acids [WO 03/097658], diglycosylated 1,2-diols [WO
97/01569], substituted 5-membered heterocycles [WO 00/33836],
mannopyranosyloxy-phenyl-benzoic acids [EP0758243 B1], piperazine
based compounds [U.S. Pat. No. 6,432,957B1], gallic acid
derivatives of peptides [WO 2004/018502], gallic acid [C. C. M.
Appeldoorn et al., Circulation 2005, 111, 106-112; EP 1481669A1],
and quinic acid derivatives [N. Kaila et al., J. Med. Chem. 2005,
48, 4346-4357 ]. However, none of these selectin-antagonizing
compounds have successfully passed clinical trials up to date [S.
J. Romano, Treat. Respir Med 2005, 4(2), 85-94; M. P. Schon,
Therapeutics and Clinical Risk Management, 2005, 1(3), 201-208].
This is due to the fact, that many of these structures have been
designed on the basis of the low potency template sLe.sup.X.
Therefore, sLe.sup.X-mimicking structures are likely to show low
potency. Other compounds show specificity against different members
of the selectin family, but antagonizing only selected selectins
can be bypassed by other selectins [M. P. Schon, Therapeutics and
Clinical Risk Management, 2005, 1(3), 201-208]. In addition, most
of the compounds developed so far have high molecular weights and
often bear carbohydrates and/or peptides making them prone to
degradation and modification by peptidases and/or glycosidases.
Carbohydrate-bearing structures have further disadvantages such as
high degree of chirality, anomericity, and low probability of
transport through lipid bilayers. Similar disadvantages arc known
for peptide-bearing compounds. Some other compounds developed for
antagonizing selectin mediated processes contain pyrogallol- and
catechol-to substructures. These motifs are prone to oxidation
processes [Kumamoto M. et al., Biosci. Biotechnol. Biochem., 2001,
65(1), 126-132] making the pharmaceutical development of these
compounds difficult. In addition, compounds with pyrogallol
substructures, such as gallic acid, are known to be cytotoxic [E.
Sergediene et al., FEBS Letters, 1999, 462, 392-396] and induce
apoptosis [K. Satoh et al., Anticancer Research, 1997, 17,
2487-2490; N. Sakaguchi et al., Biochemical Pharmacology, 1998, 55,
1973-1981].
[0011] The leading compound in the field of selectin antagonists is
bimosiamose [S. J. Romano, Treat. Respir Med 2005, 4(2), 85-94].
Presently bimosiamose [D. Bock et al., New Drugs, 2003, D04, 28,
p.28; EP 0 840 606 B1] is the most advanced compound in clinical
studies Recent investigations support the hypothesis that
bimosiamose can be considered as PSGL-1 mimetic [E. Aydt, a Wolff;
Pathobiology; 2002-2003; 70; 297-301]. This distinguishes
bimosiamose from other selectin antagonists. It is, however, a high
molecular weight compound with carbohydrate structures. The
pan-selectin antagonist bimosiamose seems to lack oral
bioavailability. Some observations indicate that bimosiamose shows
good affinity for P-selectin and a moderate affinity for E- and
L-selectin.
[0012] There is a strong medical need for novel highly potent
pan-selectin antagonists which modulate selectin-mediated function,
e.g. of selectin-dependent cell adhesion, and for the development
of methods employing such compounds to modulate conditions
associated with selectin-ligand interaction. Most of the available
anti-inflammatory pharmaceutical therapies, which are available on
the market, comprise mostly corticosteroids or NSAIDs (non
steroidal anti-inflammatory drugs) having several serious
drawbacks/side effects, and target different steps of the
inflammatory cascade. Unlike this, modulating the selectin function
is a therapeutic concept intervening the inflammation cascade at a
very early stage. Almost all promising selectin antagonists so far
failed to become marketed drugs, mostly because of low potency
and/or high molecular weight that causes problems in their
absorption-distribution-metabolism-excretion (ADME) behaviour and
thus in oral bioavailability required for the treatment of most
inflammatory disorders like rheumatoid arthritis, septic shock,
atherosclerosis, reperfusion injury and many others.
[0013] Object of the invention is to provide novel small molecules,
especially non-glycosylated/non-glycosidic and non-peptidic
compounds, which are able to potently to antagonize
selectin-mediated processes and which have less negative side
effects during their application than prior art compounds.
[0014] Unlike most of the sLe.sup.X-mimicking compounds developed
in this field, the inventive compounds are not prone to
glycosidases or peptidases. Most of the selectin antagonists
developed so far are structurally and biologically based on the
properties of sLe.sup.x or sLe.sup.a. These resulting compounds
showed, therefore, low biological activity like their template
structures. This invention, however, provides novel potent small
and drug like pan-selectin antagonists that have been invented on
the basis of biological in vitro assays mimicking PSGL-1 and
PSGL-1-like ligands or any ligands bearing sLe.sup.x or sLe.sup.a
and tyrosinesulfate motifs [N. V. Bovin; Biochem Soc Symp.;
2002;(69):143-60. N. V. Bovin; Glycoconj. J.; 1998; 15(5); 431-46.
T. V. Pochechueva et al.; Bioorg Med Chem Lett.;
2003;13(10);1709-12. G. Weitz-Schmidt et al.; Anal. Biochem.;1996;
238; 184-190].
[0015] The present invention provides pharmaceutical compositions
comprising at least one compound having the general structure of
formulas (Ia) or (Ib) and a pharmaceutically acceptable carrier
which is useful in medicine.
##STR00001##
wherein the symbols and substituents have the following meaning
--X--.dbd.
##STR00002##
[0016] with m=0,1; n=an integer from 1 to 3
##STR00003##
wherein "ring" is
##STR00004##
and with R.sup.1 being H, NO.sub.2, CP.sub.3, F, Cl, Br, I, CN,
CH.sub.3, NH.sub.2, NHAlkyl, NHAryl, NHAcyl and k=0,1
##STR00005##
T being O, S or [H,H]; p=0,1,2,
##STR00006##
the double bond is either R- or Z-configurated
##STR00007##
with -E- being --(CH.sub.2--).sub.qNH-- and q=0, 1, 2, 3
--Y.dbd.
##STR00008##
[0017] with s being 0 or 1, R.sup.2 being CO.sub.2H, CO.sub.2Alkyl,
CO.sub.2Aryl, CO.sub.2NH.sub.2, CO.sub.2Aralkyl, SO.sub.3H,
SO.sub.2NH.sub.2, PO(OH).sub.2, 1-H-tetrazolyl-, CHO, COCH.sub.3,
CH.sub.2OH, NH.sub.2, NHAlkyl, N(Alkyl)Alkyl', OCH.sub.3,
CH.sub.2OCH.sub.3, SH, F, Cl, Br, I, CH.sub.3, CH.sub.2CH.sub.3,
CN, CF.sub.3 R.sup.3 independently from R.sup.2 being H, CH.sub.3,
CH.sub.2CH.sub.3, CF.sub.3, F, Cl, Br, I, CN, NO.sub.2 and R.sup.4
independently from R.sup.2 and R.sup.3 being H, CH.sub.3,
CH.sub.2CH.sub.3, CF.sub.3, F, Cl, Br, I, CN, NO.sub.2, R.sup.2
R.sup.5 being H, NO.sub.2, CF.sub.3, F, Cl, Br, I, CN, CH.sub.3,
OCH.sub.3, SH, NH2 and --W--.dbd.--(CH.sub.2--).sub.v,
cis-CH.dbd.CH-- or trans-CH.dbd.CH--, and v being 0,1,2; in case
that --W-- is cis-CH.dbd.CH-- or trans-CH.dbd.CH--, R.sup.2 must
not be NH.sub.2 or SH;
##STR00009##
R.sup.6 independently from R.sup.2 being H, F, Cl, Me, tert-Bu,
CN,
##STR00010##
with t being 0,1,2
##STR00011##
--Z.dbd.
##STR00012##
[0019] R.sup.7 independently from R.sup.2 being H, NO.sub.2,
CF.sub.3, F, Cl, Br, I, CN, CH.sub.3, OCH.sub.3, SH, NH.sub.2,
##STR00013##
[0020] R.sup.8 independently from R.sup.2 being H, F, Cl, Me,
tert-Bu, CN, NH.sub.2
##STR00014## [0021] with K.dbd.NH, NMe, O, S
[0021] ##STR00015## [0022] or the pharmaceutically acceptable
salts, esters or amides and prodrugs of the above identified
compounds of formulas (Ia) or (Ib).
[0023] In a preferred embodiment of the invention, the compositions
comprise a compound of the formulas (Ia) or (Ib) and a
pharmaceutically acceptable carrier which is useful in a
medicine,
##STR00016##
wherein the symbols, indices and substituents have the following
meaning --X--.dbd.
##STR00017##
with m=0,1; n=an integer from 1 to 3
##STR00018##
to wherein "ring" is
##STR00019##
[0024] and with R.sup.1 being H, NO.sub.2, CF.sub.3, F, Cl, Br, I,
CN, CH.sub.3, NH.sub.2, NHAlkyl, NHAryl, NHAcyl and k=0,1
##STR00020##
T being O, S or [H,H]; p=0,1,2,
--Y.dbd.
##STR00021##
[0025] with s being 0 or 1, R.sup.2 being CO.sub.2H, CO.sub.2Alkyl,
CO.sub.2Aryl, CO2NH.sub.2, CO.sub.2Aralkyl, SO.sub.3H,
SO.sub.2NH.sub.2, PO(OH).sub.2, 1-H-tetrazolyl-, CHO, COCH.sub.3,
CH2OH, NH2, NHAlkyl, N(Alkyl)Alkyl', OCH.sub.3, CH.sub.2OCH.sub.3,
SH, F, Cl, Br, I, CH.sub.3, CH.sub.2CH.sub.3, CN, CF.sub.3 R.sup.3
independently from R.sup.2 being H, CH.sub.3, CH.sub.2CH.sub.3,
CF.sub.3, F, Cl, Br, I, CN, NO.sub.2 and R.sup.4 independently from
R.sup.2 and R.sup.3 being H, CH.sub.3, CH.sub.2CH.sub.3, CF.sub.3,
F, Cl, Br, I, CN, NO.sub.2, R.sup.2 R.sup.5 being H, NO.sub.2,
CF.sub.3, F, Cl, Br, I, CN, CH.sub.3, OCH.sub.3, SH, NH.sub.2 and
--W--.dbd.--(CH.sub.2--).sub.v, cis-CH.dbd.CH--or
trans-CH.dbd.CH--, and v being 0,1,2; in case that --W-- is
cis-CH.dbd.CH-- or trans-CH.dbd.CH--, R.sup.2 must not be NH.sub.2
or SH;
##STR00022##
with t being 0,1,2 --Z.dbd.
##STR00023##
R.sup.7 independently from R.sup.2 being H, NO.sub.2, CF.sub.3, F,
Cl, Br, I, CN, CH.sub.3, OCH.sub.3, SH, NH.sub.2,
##STR00024##
with K.dbd.NH, NMe, O, S or the pharmaceutically acceptable salts,
esters or amides and prodrugs of the above identified compounds of
formulas (Ia) or (Ib).
[0026] Preferred pharmaceutical compositions comprise compounds of
formulas (IIa) or (IIb)
##STR00025##
wherein --Y is like defined above and wherein --X'-- is X (a),
X(b), X(c), and X(d) like defined above. Preferred definitions of
--X'-- are X(a), X(b) and X(c), more preferred are X(b) and
X(c).
[0027] Further preferred pharmaceutical compositions comprise
compounds of formulas (A1), (B1), (A2) or (B2)
##STR00026##
wherein --X'-- and --Y are like defined above and wherein --X''--
is
##STR00027##
and wherein --Y' is
##STR00028##
wherein all indices, symbols and substituents are like defined
above.
[0028] In a further embodiment of the invention, the compounds of
the formulas A1,A2, B1 and B2 are used, wherein --X'-- and --Y are
as defined as above and wherein --X''-- is
##STR00029##
and wherein --Y' is
##STR00030## [0029] and wherein all other indices, symbols and
substituents are as defined above.
[0030] Particularly preferred pharmaceutical compositions comprise
compounds of formulas (C) and (D)
##STR00031##
wherein --X''-- and --Y' are like defined above.
[0031] Very particularly preferred pharmaceutical compositions
comprise at least one compound of formulas (E) and (F)
##STR00032## [0032] wherein --X''- is like defined above and --Y''
is
[0032] ##STR00033## [0033] with R.sup.9 being CO.sub.2H,
CO.sub.2alkyl, CO.sub.2aryl, CO.sub.2NH.sub.2, CO.sub.2aralkyl,
CH.sub.2SO.sub.3H, CH.sub.2SO.sub.2NH.sub.2, CH.sub.2PO(OH).sub.2,
1-H-tetrazolyl, CHO, COCH.sub.3, CH.sub.2OH, CH.sub.2NH.sub.2,
CH.sub.2NHalkyl, CH.sub.2N(alkyl)alkyl', CH.sub.2OCH.sub.3,
CH.sub.2SH, wherein the indices, symbols and substituents are
defined as above.
[0034] The invention also relates to pharmaceutical compositions,
wherein the compounds are defined by formulas (E) or (F)
##STR00034##
to wherein --X''-- is as defined as above and --Y'' is
##STR00035## [0035] with R.sup.9 being CO.sub.2H, CO.sub.2alkyl,
CO.sub.2aryl, CO.sub.2NH.sub.2, CO.sub.2aralkyl, CH.sub.2SO.sub.3H,
CH2SO.sub.2NH.sub.2, CH.sub.2PO(OH).sub.2, 1-H-tetrazolyl, CHO,
COCH.sub.3, CH.sub.2OH, CH.sub.2NH.sub.2, CH.sub.2NHalkyl,
CH.sub.2N(alkyl)alkyl', CH.sub.2OCH.sub.3, CH.sub.2SH, [0036]
wherein all other indices, symbols and substituents are as defined
above.
[0037] These chemical compounds (C), (D), (E) and (F) are also new
compounds for themselves.
[0038] All compounds as described before present the ability of
modulating cell adhesion and modulate selectin- as well as
PSGL-1-like mediated binding. The compounds have the ability to
modulate the interaction of selectins with sLe.sup.x/sLe.sup.a and
also the interaction between selectins and tyrosinesulfate
residues. Therefore they are useful for the treatment of acute and
chronic inflammatory disorders, as well as other medical conditions
where selectin mediated processes play a role.
[0039] The term "pharmaceutical" includes also diagnostic
applications.
[0040] The term "pharmaceutical" includes also prophylactic
applications in order to prevent medical conditions where selectin
mediated processes play a role.
[0041] The term "pharmaceutical" includes also applications, where
compounds of the present invention may be used as vehicles for drug
targeting of diagnostics or therapeutics.
[0042] The invention provides pharmaceutical compositions
comprising compounds of formulas (Ia) or (Ib) and in a preferred
variant of formulas (IIa) or (IIb).
[0043] In a further preferred variant the invention provides
pharmaceutical. compositions comprising at least one compound of
formula (A1), (A2), (B1) or (B2).
[0044] In a particularly preferred variant the invention provides
pharmaceutical compositions comprising at least one compound of
formula (C) or (D).
[0045] In a very particularly preferred variant the invention
provides pharmaceutical compositions comprising at least one
compound of formula (E) or (F).
[0046] The present invention further provides a method of
modulating the binding of P-selectin, L-selectin or E-selectin to
sLe.sup.x or sLe.sup.a and tyrosinesulfate residues comprising the
step of administering to a patient an effective amount of at least
one compound having the structure of formulas (Ia) or (Ib) to
modulate the binding of P-, E- or L-selectin to sLe.sup.x or
sLe.sup.a and tyrosinesulfate. It has been found that compounds
having the formulas (Ia) or (Ib) shown above act to modulate E-, P-
or L-selectin binding.
[0047] As used herein the terms "alkyl" shall mean a monovalent
straight chain or branched chain group of 1 or 2 or 3 or 4 or 5 or
6 or 7 or 8 or 9 or 10 or 11 or 12 carbon atoms including, but not
limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl,
tert-butyl and the like. "Alkyl" is independently from each other
and can be different or identical.
[0048] The term "aryl" shall mean carbocyclic and heterocyclic
aromatic groups including, but not limited to, phenyl, 1-naphthyl,
2-naphthyl, fluorenyl, (1,2)-dihydronaphthyl, indenyl, indanyl,
thienyl, benzothienyl, thienopyridyl and the like.
[0049] The term "aralkyl" (also called arylalkyl) shall mean an
aryl group appended to an alkyl group including, but not limited
to, benzyl, 1-naphthylmethyl, 2-naphthylmethyl, fluorobenzyl,
chlorobenzyl, bromobenzyl, iodobenzyl, alkoxybenzyl (wherein
"alkoxy" means methoxy, ethoxy, isopropoxy, n-butoxy, sec-butoxy,
tert-butoxy an the like), hydroxybenzyl, aminobenzyl, nitrobenzyl,
guanidinobenzyl, fluorenylmethyl, phenylmethyl(benzyl),
1-phenylethyl, 2-phenylethyl, 1-naphthylethyl and the like.
[0050] The term "acyl" shall mean --(CHO) or --(C.dbd.O)-alkyl or
--(C.dbd.O)-aryl or --(C.dbd.O)-aralkyl including, but not limited
to, formyl, acetyl, n-propionyl, isopropionyl, n-butyryl,
isobutyryl, pivaloyl, benzoyl, 4-nitrobenzoyl and the like.
[0051] The term "pharmaceutically acceptable salts, esters, amides
and prodrugs" as used herein refers to those carboxylate salts,
amino acid addition salts, esters, amides and prodrugs of the
compounds of the present invention which are, within the scope of
sound medical judgment, suitable for use in contact with tissues of
patients without undue toxicity, irritation, allergic response and
the like, commensurate with a reasonable benefit/risk ratio, and
effective for their intended use, as well as the zwitterionic
forms, where possible, of the compounds of the present invention.
The term "salts" refers to the relatively non-toxic, inorganic and
organic acid addition salts of the compounds of the present
invention. These to salts can be prepared in situ during the final
isolation and purification of the compounds or by separately
reacting the purified compounds in its free form with a suitable
inorganic or organic acid or base and isolating the salt thus
formed. Representative salts of the compounds of the present
invention include the hydrobromide, hydrochloride, sulfate,
bisulfate, nitrate, acetate, oxalate, valerate, palmitate,
stearate, laurate, borate, benzoate, lactate, phosphate, tosylate,
citrate, maleate, fumarate, succinate, tartrate, naphthylate,
mesylate, glucoheptonate, lactiobionate, laurylsulphonate salts and
the like. These may include cations based on the alkali and
alkalineearth metals, such as sodium, lithium, potassium, calcium,
magnesium and the like, as well as non-toxic ammonium, quaternary
ammonium and amine cations including, but not limited to, ammonium,
tetramethylammonium, tetraethylammoni urn, methylamine,
dimethylamine, trimethylamine, triethylamine, ethylamine, and the
like.
[0052] Examples of the pharmaceutically acceptable, non-toxic
esters of the compounds of this invention include C.sub.1, C.sub.2,
C.sub.3, C.sub.4, C.sub.5 and C.sub.6 alkyl esters wherein the
alkyl group is a straight or branched chain. Acceptable esters also
include C.sub.5, C.sub.6 and C.sub.7 cycloalkyl esters as well
arylalkyl esters such as, but not limited to benzyl. C.sub.1,
C.sub.2, C.sub.3, C4, C.sub.5 and C.sub.6 alkyl ester are
preferred. Esters of the compounds of the present invention may be
prepared according to conventional methods.
[0053] Examples of pharmaceutically acceptable, non-toxic amides of
compounds of this invention include amides derived from ammonia,
primary C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5 and C.sub.6
alkyl amines and secondary C.sub.1, C.sub.2, C.sub.3, C.sub.4,
C.sub.5 and C.sub.6 dialkyl amines wherein the alkyl groups are
straight or branched chains. In the case of secondary amines the
amine may also be in the form of a 5 or 6 membered heterocycle
containing one nitrogen atom. Amides derived from ammonia, C.sub.1,
C.sub.2 and C.sub.3 alkyl primary amides and C.sub.1 to C.sub.2
dialkyl secondary amides are preferred. Amides of the compounds of
the present invention may be prepared according to conventional
methods.
[0054] The term "prodrug" refers to one or more compounds that are
rapidly transformed in vitro and from a non-active to active state
in vivo to yield the parent compound of the above formulas (Ia) or
(Ib), for example by hydrolysis in blood or in vivo metabolism.
[0055] It is also contemplated that pharmaceutically active
compositions may contain a compound of the present invention or
other compounds that modulate or compete with E-selectin or
P-selectin or L-selectin binding.
[0056] Pharmaceutically active compositions of the present
invention comprise a pharmaceutically acceptable carrier and a
compound of formulas (Ia) or (Ib), whereby a pharmaceutically
acceptable carrier can also be a medically appropriate
nano-particle, dendrimer, liposome, microbubble or polyethylene
glycol (PEG). The pharmaceutical compositions of the present
invention may include one or more of the compounds having the above
structure (Ia) or (Ib) formulated together with one or more,
physiologically acceptable carriers, adjuvants or vehicles, which
are collectively referred to herein as carriers, for parenteral
injection, for oral administration in solid or liquid form, for
rectal or topical administration and the like.
[0057] The compositions can be administered to humans and animals
either orally, rectally, parenterally (intravenously,
intramuscularly, intradermaly or subcutaneously), intracisternally,
intravaginally, interperitoneally, locally (powders, ointments or
drops), or as a buccal or by inhalation (nebulized, or as nasal
sprays).
[0058] Compositions suitable for parenteral injection may comprise
physiologically acceptable sterile aqueous or nonaqueous solutions,
stabilizers, antioxidants, preservatives (e.g. ascorbic acid,
sodium sulfite, sodium hydrogene sulfite, benzyl alcohol, EDTA),
dispersions, suspensions or emulsions and sterile powders for
reconstitution into sterile injectable solution or dispersion.
Examples of suitable aqueous and nonaqueous carriers, diluents,
solvents or vehicles include water, ethanol, polyol, (propylene
glycol, polyethylene glycol, glycerol and the like), suitable
mixtures thereof, vegetable oils (such as olive or canola oil) and
injectable organic esters such as ethyl oleate. Proper fluidity can
be maintained, for examples, by the use of a coating such as
lecithin, by the maintenance of the required particle size in the
case of dispersions and by the use of surfactants.
[0059] These compositions may also contain adjuvants such as
preserving, wetting, emulsifying, and dispersing agents. Prevention
of the actions of microorganisms can be ensured by various
antibacterial and antifungal agents, for examples, parabens,
chlorobutanol, phenol, sorbic acid, and the like. It may also be
desirable to include isotonic agents, for examples sugars, sodium
chloride and the like. Prolonged absorption of the injectable
pharmaceutical form can be brought about by the use of agents
delaying absorption, for examples aluminium monostearate and
gelatin.
[0060] If desired, and for more effective distribution, the
compounds can be incorporated into slow or timed release or
targeted delivery systems such as polymer matrices, liposomes, and
microspheres. They may be sterilized, for example, by filtration
through a bacteria-retaining filter, or by incorporating
sterilizing agents in the form of sterile water, or some other
sterile injectable medium immediately before use.
[0061] Solid dosage forms for oral administration include capsules,
tablets, pills, powders and granules. In such solid dosage forms,
the active compound or a prodrug is admixed with at least one inert
customary excipient (or carrier) such as sodium citrate or
dicalcium phosphate or (i) fillers or extenders, as for example,
starches, lactose, sucrose, glucose, mannitol and silicic acid,
(ii) binders, as for example, carboxymethylcellulose, alginates,
gelatine, polyvinylpyrrolidone, sucrose and acacia, (iii)
humectants, as for example, glycerol, (div disintegrating agents,
as for example, agar-agar, calcium carbonate, potato or tapioca
starch, aliginic acid, certain complex silicates and sodium
carbonate, (v) solution retarders, as for examples, paraffin, (vi)
absorption accelerators, as for example, quaternary ammonium
compounds, (vii) wetting agents, as for examples, cetyl alcohol and
glycerol monostearate, (viii) adsorbents, as for example, kaolin
and bentonite, and (ix) lubricants, as for example, talc, calcium
stearate, magnesium stearate, solid polyethylene glycols, sodium
lauryl sulfate and mixtures thereof. In the case of capsules,
tablets, and pills, the dosage forms may also comprise buffering
agents.
[0062] Solid compositions of a similar type may also, be employed
as fillers in soft and hard-filled gelatine capsules using
excipients as lactose or milk sugars as well as high molecular
polyethylene glycols and the like. Solid dosage forms such as
tablets, dragees, capsules, pills and granules can be prepared with
coatings and shells, such as enteric coatings and others well known
in the art. They may contain opacifying agents, and can also be of
such compositions that they release the active compound or
compounds in a certain part of the intestinal tract in a delayed
manner. Examples of embedding compositions that can be used are
polymeric substances and waxes. The active compounds can also be in
microencapsulated form, if appropriate, with one or more of the
above-mentioned excipients.
[0063] Liquid dosage forms for oral administration include
pharmaceutically acceptable emulsions, solutions, suspensions,
syrups and elixirs. In addition to the active compounds, the liquid
dosage forms may contain inert diluents commonly used in the art
such as water or other solvents, solubilizing agents and
emulsifiers, as for example, ethyl alcohol, isopropyl alcohol,
ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, in
particular, cottonseed oil, groundnut oil, corn germ oil, olive
oil, cannola oil, caster oil and sesame seed oil, glycerol,
tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid
esters of sorbitan or mixtures of these substances, and the like.
Besides such inert diluents, the compositions can also include
adjuvants, such as wetting agents, emulsifying and suspending
agents, sweeting, flavouring and perfuming agents.
[0064] Suspensions, in addition to the active compounds, may
contain suspending agents, for example, ethoxylated isostearyl
alcohols, polyoxyethylene sorbitol and sorbitan esters,
microcrystalline cellulose, aluminium metahydroxide, bentonite,
agar-agar, tragacanth or mixtures of these substances and the
like.
[0065] Compositions for rectal administrations are preferably
suppositories, which can be prepared by mixing the compounds of the
present invention with suitable nonirritating excipients or
carriers such as cacao butter, polyethylene glycol or a suppository
wax, which are solid at ordinary temperatures but liquid at body
temperature and therefore melt in the rectal or vaginal cavity and
release the active component. Dosage forms for topical
administration of a compound of this invention include ointments,
powder, sprays and inhalants.
[0066] The active component is admixed under sterile conditions
with a physiologically acceptable carrier and any needed
preservatives, buffers or propellants as may be required.
Ophthalmic formulations, eye ointments, suspensions, powder and
solutions are also contemplated as being within the scope of this
invention.
[0067] The compounds of the present invention can also be
incorporated into or connected to liposomes or administrated in the
form of liposomes. As is known in the art, liposomes are generally
derived from phospholipids or other lipid substances. Liposomes are
formed by mono or multilamellar hydrated liquid crystals that are
dispersed in an aqueous medium. Any non-toxic, physiologically
acceptable metabolized lipid capable of forming liposomes can be
used. The present compositions in liposome form can contain, in
addition to the selectin binding antagonists of the present
invention, stabilizers, preservatives, excipients and the like. The
preferred lipids are the phospholipids and the phosphatidyl
cholines (lecithins), both natural and synthetic. Methods to form
liposomes are well known in the art.
[0068] Non-parenteral dosage forms may also contain a
bioavailability enhancing agent (e.g. enzyme modulators,
antioxidants) appropriate for the protection of the compounds
against degradation. Actual dosage levels of active ingredient in
the composition of the present invention may be varied so as to
obtain an amount of active ingredient that is effective to obtain
the desired therapeutic response for a particular composition and
method of administration. The selected dosage level, therefore,
depends on the desired therapeutic effect, on the route of
administration, on the desired duration of treatment and other
factors. The total daily dosage of the compounds on this invention
administered to a host in to single or divided doses may be in the
range up to 50 mg per kilogram of body weight. Dosage unit
compositions may contain such submultiples thereof as may be used
to make up the daily dosage. It will be understood, however, that
the specific dose level for any particular patient, whether human
or other animal, will depend upon a variety of factors including
the body weight, general health, sex diet, time and route of
administration, rates of absorption and excretion, combination with
other drugs and the severity of the particular disease being
treated.
[0069] In particular, the compounds of the present invention may be
used to treat a variety of diseases relating to inflammation and
cell-cell recognition and adhesion. For example, the compounds of
the present invention may be administrated to a patient to treat
Chronic Obstructive Pulmonary Disease (COPD), acute lung injury
(ALI), cardiopulmonary bypass, acute respiratory distress syndrome
(ARDS), Crohn's disease, septic shock, sepsis, chronic inflammatory
diseases such as psoriasis, atopic dermatitis, and rheumatoid
arthritis, and reperfusion injury that occurs following heart
attacks, strokes, atherosclerosis, and organ transplants, traumatic
shock, multi-organ failure, autoimmune diseases like multiple
sclerosis, percutaneous transluminal angioplasty, asthma and
inflammatory bowel disease. In each case, an effective amount of
the compounds of the present invention is administered either alone
or as part of a pharmaceutically active composition to a patient in
need of such treatment. It is also recognized that a combination of
the compounds may be administered to a patient in need of such
administration. The compounds of the present invention may also be
administered to treat other diseases that are associated with
cell-cell adhesion. As the present compounds modulate the binding
of E-selectin or P-selectin or L-selectin, any disease that is
related to this interaction may potentially be treated by the
modulation of this binding interaction.
[0070] In addition to being found on some white blood cells,
sLe.sup.a is found on various cancer cells, including lung and
colon cancer cells. It has been suggested that cell adhesion
involving sLe.sup.a may be involved in the metastasis of certain
cancers and antagonists of sLe.sup.a binding might be useful in
treatment of some forms of cancer.
[0071] The use of the active ingredients according to the invention
or of cosmetic or topical dermatological compositions with an
effective content of active ingredient according to the invention
surprisingly enables effective treatment, but also prophylaxis of
skin ageing caused by extrinsic and intrinsic factors.
[0072] The invention particularly relates to the use of a compound
of formula (Ia) or (Ib) or a stereoisomeric form thereof for the
preparation of a cosmetic or dermatological composition.
[0073] The amount used of the active compound or a stereoisomeric
form thereof corresponds to the amount required to obtain the
desired result using the cosmetic or dermatological compositions.
One skilled in this art is capable of evaluating this effective
amount, which depends on the derivative used, the individual on
whom it is applied, and the time of this application. To provide an
order of magnitude, in the cosmetic or dermatological compositions
according to the invention, the compound of formula (Ia) or (Ib) or
a stereoisomeric form thereof may be administered in an amount
representing from 0.001% to 40% by weight, preferentially 0.005% to
30% by weight and more preferentially from 0.01% to 20% by
weight.
[0074] A further aspect covers cosmetic compositions comprising a
compound of formula (Ia) or (Ib) or a stereoisomeric form thereof
and at least one cosmetically tolerable component, e.g. a
cosmetically tolerable component for skin applications.
[0075] The amounts of the various components of the physiological
medium of the cosmetic composition according to the invention are
those generally included in the fields under consideration. When
the cosmetic composition is an emulsion, the proportion of the
fatty phase may range from 2% to 80% by weight and preferably from
5% to 50% by weight relative to the total weight of the cosmetic
composition.
[0076] Thus, the cosmetic composition should contain a non-toxic
physiologically acceptable medium that can be applied to human
skin. For a topical application to the skin, the cosmetic
composition may be in the form of a solution, a suspension, an
emulsion or a dispersion of more or less fluid consistency and
especially liquid or semi-liquid consistency, obtained by
dispersing a fatty phase in an aqueous phase (O/W) or, conversely,
(W/O), or alternatively a gel. A cosmetic composition in the form
of a mousse or in the form of a spray or an aerosol then comprising
a pressurized propellant may also be provided. Also the
compositions may be in the form of a haircare lotion, a shampoo or
hair conditioner, a liquid or solid soap, a treating mask, or a
foaming cream or gel for cleansing the hair. They may also be in
the form of hair dye or hair mascara.
[0077] The cosmetic compositions of the invention may also comprise
one or more other ingredients usually employed in the fields under
consideration, selected from among formulation additives, for
instance aqueous-phase or oily-phase thickeners or gelling agents,
dyestuffs that are soluble in the medium of the cosmetic
composition, solid particles such as mineral or organic fillers or
pigments in the form of microparticles or nanoparticles,
preservatives, fragrances, hydrotopes or electrolytes, neutralizers
(acidifying or basifying agents), propellants, anionic, cationic or
amphoteric surfactants, polymers, in particular water-soluble or
water-dispersible anionic, nonionic, cationic or amphoteric
film-forming polymers, mineral or organic salts, chelating agents;
mixtures thereof.
[0078] The cosmetic compositions may be used to inhibit the
micro-inflammatory cycle. Thus, the present invention also relates
to cosmetic compositions comprising a compound of formula (Ia) or
(Ib) or a stereoisomeric form thereof that is used for the cosmetic
treatment or cosmetic prophylaxis of micro-inflammatory
conditions.
[0079] Cosmetic compositions comprising a compound of formula (Ia)
or (Ib) or a stereoisomeric form thereof that is used for the
cosmetic treatment or cosmetic prophylaxis of skin ageing caused by
intrinsic factors are also subject of the present invention.
Intrinsic factors responsible for skin ageing are genetically
programmed determinants including age, hormonal status, and
psychological factors.
[0080] Beside cosmetically inactive ingredients the cosmetic
compositions of the present invention may also comprise one or more
cosmetically active ingredients with beneficial to action on the
skin. Thus, the present invention relates to cosmetic compositions
comprising a compound of formula (Ia) or (Ib) or a stereo isomeric
form thereof and at least one further cosmetically active
ingredient, e.g. an UV-blocker or proteins.
[0081] Dermatological compositions comprising a compound of formula
(Ia) or (Ib) or a stereoisomeric form thereof and at least one
dermatologically tolerable component, e.g. a dermatologically
tolerable component for skin applications, are also subject of the
invention.
[0082] Dermatologically tolerable components that can be used for
the dermatological compositions described here are identical to the
cosmetically tolerable components as defined in this invention.
[0083] A further embodiment of this invention are dermatological
compositions comprising a compound of formula (Ia) or (Ib) or a
stereoisomeric form thereof that is used for the dermatological
treatment, dermatological diagnosis or dermatological prophylaxis
of micro-inflammatory conditions.
[0084] In particular the invention covers dermatological
compositions comprising a compound of formula (Ia) or (Ib) or a
stereoisomeric form thereof that is used for the dermatological
treatment, dermatological diagnosis or dermatological prophylaxis
of itching and skin ageing caused by extrinsic factors. Extrinsic
factors include environmental factors in general; more particularly
photo-ageing due to exposure to the sun, to light or to any other
radiation, atmospheric pollution, wounds, infections, traumatisms,
anoxia, cigarette smoke, hormonal status as response to external
factors, neuropeptides, electromagnetic fields, gravity, lifestyle
(e.g. excessive consumption of alcohol), repetitive facial
expressions, sleeping positions, and psychological stressors.
[0085] In addition to dermatologically inactive ingredients the
dermatological compositions may also comprise dermatologically or
pharmaceutically active ingredients. Thus, the present invention
also relates to dermatological compositions comprising a compound
of formula (Ia) or (Ib) or a stereoisomeric form thereof and at
least one further dermatologically or pharmaceutically active
ingredient. The dermatologically or pharmaceutically active
ingredients that can be used for the dermatological compositions
described herein are defined as the cosmetically active ingredients
defined above. Dermatologically or pharmaceutically active
ingredients can be identical to the cosmetically active ingredients
as defined in this invention.
[0086] Another subject of the present invention are dermatological
compositions comprising a compound of formula (Ia) or (Ib) or a
stereoisomeric form thereof and at least one further
dermatologically or pharmaceutically active ingredient
characterized in that it is used for the dermatological treatment,
dermatological diagnosis or dermatological prophylaxis of
micro-inflammatory conditions.
[0087] In particular, the present invention relates to
dermatological compositions comprising a compound of formula (Ia)
or (Ib) or a stereoisomeric form thereof and at least one further
dermatologically or pharmaceutically active ingredient
characterized in that it is used for the dermatological treatment,
dermatological diagnosis or dermatological prophylaxis of itching
and skin ageing caused by extrinsic factors.
[0088] Ageing of the skin may also be caused by a combination of
intrinsic and extrinsic factors. Therefore, the present invention
also relates to dermatological compositions comprising a compound
of formula (Ia) or (Ib) or a stereoisomeric form thereof and at
least one further pharmaceutically or cosmetically active
ingredient characterized in that it is used for the cosmetic and
dermatological treatment and cosmetic and dermatological
prophylaxis of skin ageing caused by a combination of intrinsic and
extrinsic factors.
[0089] Another embodiment of this invention is a process for the
preparation of a cosmetic composition by mixing a compound of
formula (Ia) or (Ib) or a stereoisomeric form thereof, at least one
cosmetically tolerable component and eventually further
cosmetically active ingredients.
[0090] In particular, a process for the preparation of a cosmetic
composition by mixing a compound of formula (Ia) or (Ib) or a
stereoisomeric form thereof, at least one cosmetically tolerable
component and eventually further cosmetically active ingredients,
wherein the composition includes from 0.01% to 20% by weight of
compound of formula (Ia) or Ib) or a stereoisomeric form thereof,
based on the total weight of the composition is subject of this
invention.
[0091] A further aspect deals with a process for the preparation of
a dermatological composition by mixing a compound of formula (Ia)
or (Ib) or a stereoisomeric form thereof, at least one
dermatologically tolerable component and eventually further
pharmaceutically active ingredients.
[0092] Many of the compounds of the present invention may be
synthesized according to the following general synthetic
schemes.
##STR00036##
[0093] In SCHEME 1 an amino acid of type (1) is reacted to the
corresponding methyl ester (2) under heating with acidic methanol.
Ester (2) is reacted with a trimethoxy-phenyl-alkylic acid under
state-of-the-art conditions (i.e.
N'-(3-dimethylaminopropyl)-N-ethyl carbodiimide (EDC),
triethylamine and 4-dimethylaminopyridine (DMAP) in a chlorinated
solvent) to the amide (3). Alternatively diisopropyl carbodiimide
(DIC) and hydroxybenzotriazole (HOBt) may be used for this reaction
step. The synthesis sequence shown in SCHEME 1 leading to compounds
like (3) is not only reduced to the Y-H building blocks like (1)
but may be generally applied to all other Y-H type building blocks
leading to compounds of type (A1), (A2), (B1) and (B2) as shown in
the paragraph before.
##STR00037##
[0094] In SCHEME 2 a carboxy substituted thiophene like (4) is
reacted to the corresponding ethyl. ester (5) under heating in
acidic ethanol. Ester (5) is brominated with N-bromosuccinimide in
anhydrous chloroform and glacial acetic acid to give (6) which is
further reacted with 2-Amino-benzeneboronic acid under a
state-of-the-art Suzuki transformation (i.e.
Tetrakis(triphenylphosphine)-palladium, aqueous sodium carbonate,
ethanol, toluene) to the biaryl (7). Biaryl (7) is reacted with a
trimethoxy-phenyl-alkylic acid, EDC, triethylamine and DMAP in a
chlorinated solvent to the amide (8). Alternatively DIC and HOBt
may be used for this reaction step.
##STR00038##
[0095] In SCHEME 3 Methyl-3-bromobenzoate (9) is reacted under
inert conditions with a Trimethoxyphenylboronic acid under
Suzuki-type basic conditions (Pd(PPh.sub.3).sub.4 and aqueous
sodium bicarbonate in dimethoxyethane) to a biphenyl of type (10)
which is further hydrolized with aqueous lithium hydroxide in
acetonitrile to give the corresponding carboxylic acid (11) which
was converted to building block of type (12) by reaction with
oxalyl chloride in anhydrous dichloromethane.
##STR00039##
[0096] In SCHEME 4 an acid chloride like (12) is reacted with an
aniline of general type (13) under basic conditions (triethylamine
in a chlorinated solvent) to form the anilide (14). Alternatively
pyridine may be used for this reaction step.
[0097] In case that R.sup.2 and/or R.sup.4 contain carboxylic acid
functionalities, those are protected as their corresponding methyl
or ethyl esters before and hydrolized afterwards to release the
carboxylic acid functionalities. The ester hydrolysis is done with
LiOH in MeCN or THF/MeOH.
[0098] The synthesis sequence shown in SCHEME 4 leading to
compounds like (14) is not only reduced to X--Y--H and Y--H
building blocks like (13) but may be generally applied to all other
X--Y--H and Y--H type building blocks leading to compounds of type
(A1), (A2), (B1) and (B2) as shown in the paragraphs before.
##STR00040##
[0099] In SCHEME 5 the generation of building block (19) is
outlined, whereby the furan (16) is available by NBS-bromination of
methyl furoate (15) and pinacolyl borane of type (18) is available
by Pd-catalyzed boration of anilines like (17). Suzuki-type
coupling of (16) and (18) with Pd(PPh.sub.3).sub.4 leads to biaryls
of type (19).
##STR00041##
In SCHEME 6 a biaryl of type (19) is reacted with a
trimethoxy-phenyl-alkylic acid under state-of-the-art conditions
(i.e. N'-(3-dimethylaminopropyl)-N-ethyl carbodiimide (EDC),
triethylamine and 4-dimethylaminopyridine (DMAP) in a chlorinated
solvent) to the amide of type (20). Alternatively diisopropyl
carbodiimide (DIC) and hydroxybenzotriazole (HOBt) may be used for
this reaction step. (20) is then hydrolized to acid of type (21)
whether with LiOH in MeCN or THF/MeOH.
[0100] The present invention is furthermore illustrated by the
following representative examples.
EXAMPLE 1
{3-[3-(2,3,4-Trimethoxy-phenyl)-propionylamino]-phenyl}-acetic acid
methyl ester (24)
##STR00042##
[0102] Step 1:Dissolve (3-Amino-phenyl)-acetic acid ((22), 700 mg,
4.63 mmol) in MeOH (21 mL) and add conc. sulfuric acid (0.27 mL,
5.09 mmol). Stir the reaction mixture for 2 d under reflux. Cooled
mixture to room temperature (rt), remove solvent under reduced
pressure and prepurify the residue by flushing it over a short pad
of silica gel using EtOAc. Remove solvent again and partition the
residue between EtOAc and saturated aqu. NaHCO.sub.3 (1+1).
Extracte the aqueous layer 3 times with EtOAc, washe the combined
organic layers with brine and dried with Na.sub.2SO.sub.4. Remove
solvent under reduced pressure and dry the residue without further
purification in oil pump vacuum to obtain product (23) as a light
yellow oil (708 mg, 92%). NMR (400 MHz, CDCl.sub.3): 3.51 (s, 2 H);
3.67 (s, 3 H); 6.57 (dd, 1 H, J.sub.1=7.8 Hz, J.sub.2=1.8 Hz); 6.60
(br..PSI.t, 1 H, J=1.8 Hz); 6.65 (br.d, 1 H, J.apprxeq.7.8 Hz);
7.08 (.PSI.t, 1 H, J=7.8 Hz).
[0103] Step 2: (The following reaction is done in an anhydrous
N.sub.2 atmosphere.) Dissolve EDC hydrochloride (187 mg, 0.98 mmol)
and triethylamine (0.14 mL, 1.00 mmol) in anhydrous dichloromethane
(3.5 mL) and stir for 5 min at rt. Added
3-(2,3,4-Trimethoxy-phenyl)-propionic acid (234 mg, 0.97 mmol) and
DMAP (12 mg, 0.10 mmol) and stir for 10 min. Add ester (23) (107
mg, 0.65 mmol) and stir the reaction solution overnight at rt.
[0104] Hydrolize the reaction solution with saturated aqu.
NH.sub.4Cl followed by water, separate layers, extracte aqu. layer
with dichloromethane (3 times) and washe the combined organic
layers with water and brine and dry with Na.sub.2SO.sub.4. Remove
solvent under reduced pressure.
[0105] Purify crude product by preparative radial chromatography
(silica gel 60PF, EtOAc/CyH 1+1) to obtain product (24) as a white
solid (209 mg, 83%). [K. C. Nicolaou; P. S. Baran; Y.-L. Thong; K.
Sugita; J. Am. Chem. Soc.; 2002; 124; 10; 2212-2220]. .sup.1H NMR
(400 MHz, CDCl.sub.3): 2.62 (t, 2 H, J=7.5 Hz); 2.95 (t, 2 H, J=7.5
Hz); 3.58 (s, 2 H); 3.67 (s, 3 H); 3.82 (s, 3 H); 3.84 (s, 3 H);
3.91 (s, 3 H); 6.59 (d, 1 H, J=8.6 Hz); 6.86. (d, 1 H, J=8.6 Hz);
6.98 (br.d, 1 H, J=7.8 Hz); 7.32 (.PSI.t, 1 H, J=7.8 Hz); 7.38
(br.d, 1 H, J=7.8 Hz); 7.41 (br.s, 1 H).
EXAMPLE 2
(5-{2-[2-(2,3,4-Trimethoxyphenyl)-acetylamino]-phenyl}-thiophen-2-yl]aceti-
c acid (31)
##STR00043## ##STR00044##
[0107] Step 1: Dissolve Thiophene-2-yl-acetic acid (25) (2.44 g,
17.1 mmol) in ethanol (35 mL) and add fuming aqu. hydrochloric acid
(few drops). Stir the reaction mixture for 19 h at 70 .degree. C.
Cool mixture to rt, remove solvent under reduced pressure and
resolve the residue in EtOAc. Wash this organic layer 3 times with
5% aqu. Na.sub.2CO.sub.3 and extract the combined aqueous layer 3
times with EtOAc. Wash the combined organic layers with brine and
dry with Na.sub.2SO.sub.4. Remove solvent under reduced pressure
and dry the residue without further purification in oil pump vacuum
to obtain product (26) as a light brown oil (2.78 g, 95%). [J.
Kunes; V. Balsanek; M. Pour; V. Buchta; Collect. Czech. Chem.
Commun., 2001, 66; 12; 1809-1830]. NMR (400 MHz, CDCl.sub.3): 1.26
(t, 3 H, J=7.1 Hz); 3.81 (s, 2 H); 4.17 (q, 2 H, J=7.1 Hz);
6.91-6.96 (m, 2 H); 7.20 (d, 1 H, J=4.8 Hz).
[0108] Step 2: (The following reaction is done in an anhydrous
N.sub.2 atmosphere.) Dissolve ester (26) (1.30 g, 7.64 mmol) in
anhydrous chloroform (6.0 mL) and glacial acetic acid (6.0 mL), add
N-Bromosuccinimide (1.39 g, 7.79 mmol) in portions and stir the
mixture for 23 h at rt. The mixture is diluted with an equal volume
of water, the organic layer separated and washed with a 1M aqu.
NaOH, water, again with 1M aqu. NaOH and water (2 times). Finally
wash the organic layer with brine and dry with Na.sub.2SO.sub.4.
Remove solvent under reduced pressure. Purify crude product by
preparative radial chromatography (silica gel 60PF, CyH/EtOAc 5+1]
to obtain product (27) as an impured (according to NMR: 20%
sideproduct) orange liquid (1.61 g, 85%) which is used without any
further purification. [P. M. Jackson; C. J. Moody; P. Sha; J. Chem.
Soc. Perkin Trans. 1; 1990; 2909-2918]. .sup.1H NMR (400 MHz,
CDCl.sub.3): 1.26 (t, 3 H, J=7.1 Hz), 3.73 (s, 2 H); 4.17 (q, 2 H,
J=7.1 Hz); 6.67 (d, 1 H, J=3.5 Hz); 6.88 (d, 1 H, J=3.5 Hz).
[0109] Step 3: (The following reaction is done in an oxygenfree
N.sub.2 atmosphere.) Ethanol (1.47 mL),
Tetrakis-(triphenylphosphine)-palladium(0) (59.0 mg, 2.5mol %) and
aqu. Na.sub.2CO.sub.3 (1.60 g, 5.60 mmol; presolved in 2.0 mL
H.sub.2O) are subsequently added to dissolved
2-Amino-benzeneboronic acid (28) (341 mg, 2.20 mmol) in toluene (16
mL). The reaction mixture is degassed 5 times and flooded with
N.sub.2 again. Add bromide (27) (498 mg, 2.00 mmol) and rinse with
toluene (4.5 mL), degas again (5 times) and stir the reaction
solution 21 h at 100.degree. C. Partition the reaction solution
between EtOAc and brine (1+1) and extract the separated aqueous
layer 3 times with EtOAc. Wash combined organic layer with brine
and dry with Na.sub.2SO.sub.4. Remove solvent under reduced
pressure and purify the crude product by preparative radial
chromatography (silica gel 60PF, CyH/EtOAc 6+1, later 3+1] to
obtain product (29) as a light yellow solid (300 mg, 57%). [N.
Miyaura; A. Suzuki; Chem. Rev.; 1995; 95; 2457]. .sup.1H NMR (400
MHz, CDCl.sub.3): 1.28 (t, 3 H, J=7.1 Hz); 3.82 (s, 2 H); 4.19 (q,
2 H, J=7.1 Hz); 6.77-6.84 (m, 2 H); 6.91 (d, 1 H, J=3.5 Hz); 7.04
(d, 1 H, J=3.5 Hz); 7.13 (td, 1 H, J=7.8 Hz, 1.3 Hz); 7.25 (d, 1 H,
J=7.8 Hz).
[0110] Step 4: (The following reaction is done in an anhydrous
N.sub.2 atmosphere.) Suspend EDC hydrochloride (86.3 mg, 0.45 mmol)
in anhydrous dichloromethane (1.4 mL), add triethylamine (0.063 mL,
0.45 mmol) and stir for 10 min at rt. Add
2-(2,3,4-Trimethoxy-phenyl)-acetic acid (74.7 mg, 0.33 mmol) and
DMAP (3.7 mg, 0.03 mmol) and stir for 15 min. Added ester (29)
(64,9 mg, 0.30 mmol) and stir the reaction solution 22 h at rt.
Partition the reaction solution between dichloromethane and water
(1+1), separate layers and extract aqu. layer with dichloromethane
(3 times). Wash the combined organic layer with brine and dry with
Na.sub.2SO.sub.4. Purify crude product by preparative radial
chromatography (silica gel 60PF, CyH/EtOAc 3+2) to obtain product
(30) as yellow oil (118 mg, 84%). .sup.1H NMR (400 MHz,
CDCl.sub.3): 1.29 (t, 3 H, J=7.1 Hz); 3.58 (s, 2 H); 3.74 (s, 3 H);
3.78 (s, 3 H); 3.79-3.80 (m, 2 H); 3.86 (s, 2 H); 4.20 (q, 2 H,
J=7.1 Hz); 6.58 (d, 1 H, J=8.6 Hz); 6.59 (d, 1 H, J=3.5 Hz); 6.75
(d, 1 H, J=3.5 Hz); 6.85 (d, 1 H, J=8.6 Hz); 7.05 (t, 1 H, J=7.8
Hz); 7.26 (dd, 1 H, J=7.8 Hz, 1.3 Hz); 7.30 (td, 1 H, J=7.8 Hz, 1.3
Hz); 7.90 (br.s; 1 H), 8.38 (d, 1 H, J=8.3 Hz).
[0111] Step 5: Dissolve ester (30) (118 mg, 0.25 mmol) in methanol
(8.0 mL), add a 1M aqu. LiOH solution (1.76 mL, 1.76 mmol) and stir
20 h at rt. Remove solvent under reduced pressure and partition
residue between CHCl.sub.3 and 0.5 M HCl (1+1). Separate the
aqueous layer and extract 3 times with CHCl.sub.3. Wash the
combined organic layer with brine and dry with Na.sub.2SO.sub.4.
Remove solvent under reduced pressure and dry the residue without
further purification in oil pump vacuum to obtain crude product
(31) as light brown foam (120 mg, quant.). NMR (400 MHz,
CDCl.sub.3): 3.58 (s, 2 H); 3.73 (s, 3 H); 3.78 (s, 3 H); 3.85 (s,
2 H); 3.86 (s, 3 H); 6.58-6.61 (m, 1 H); 6.59 (d, 1 H, J=8.3 Hz);
6.77 (d, 1 H, J=3.5 Hz); 6.86 (d, 1 H, J=8.3 Hz); 7.06 (t, 1 H,
J=7.8 Hz); 7.22-7.27 (m, 1 H); 7.31 (td, 1 H, J=7.8 Hz, 1.3 Hz);
7.86 (br.s, 1 H); 8.37 (d, 1 H, J=8.3 Hz).
EXAMPLE 3
(5-{2-[(2',3',4'-Trimethoxy-biphenyl-3-carbonyl)-amino]-phenyl}-thiophen-2-
-yl)-acetic acid methyl ester (37)
##STR00045##
[0113] Step 1: (The following reaction is done in an N.sub.2
atmosphere.) To a solution of 2,3,4-Trimethoxyphenylboronic acid
(32) (1.40 g, 6.60 mmol) in toluene (15.0 mL) is added EtOH (2.0
mL), Pd(PPh.sub.3).sub.4 (208 mg, 0.18 mmol) and Na.sub.2CO.sub.310
H.sub.2O (4.81 g, 16.80 mmol) in water (5.2 mL). The resulting
mixture is carefully degassed (5 times alternating vacuum and
flushing with N.sub.2). A solution of Methyl-3-bromobenzoate (9)
(1.29 g, 6.00 mmol) in toluene (9.0 mL) is added by syringe, the
resulting mixture is again carefully degassed and stirred overnight
at 100.degree. C. Partition the mixture between brine/EtOAc (1+1),
separate layers, extract the aqu. layer with EtOAc (3.times.), wash
the combined organic layer with brine, dry with Na.sub.2SO.sub.4
and remove solvent. Purify crude product by preparative radial
chromatography (silica gel, EtOAc/CyH 1+5) to obtain
2',3',4'-Trimethoxy-biphenyl-3-carboxylic acid methyl ester (33) as
a yellowish oil (1.07 g, 58%). .sup.1H NMR (400 MHz, CDCl.sub.3):
3.66 (s, 3 H); 3.89 (s, 3 H); 3.92 (s, 6 H); 6.74 (d, 1 H, J=8.6
Hz); 7.03 (d, 1 H, J =8.6 Hz); 7.44 (t, 1 H, J=7.8 Hz); 7.70 (d, 1
H, J=7.6 Hz); 7.97 (d, 1 H, J=7.8 Hz); 8.15 (br.s 1 H).
[0114] Step 2: Dissolve 2',3',4'-Trimethoxy-biphenyl-3-carboxylic
acid methyl ester (33) (566 mg, 1.87 mmol) in MeCN (19.0 mL) at rt
and add 1M aqu LiOH (9.36 mL, 9.36 mmol). Stir reaction mixture
overnight at rt. Quench reaction mixture (cooling bath) with 1M
aqu. HCl (to get pH ca. 3). Extract the mixture with EtOAc
(3.times.), wash the combined organic layer with brine and dry with
Na.sub.2SO.sub.4. Recrystallize crude product from EtOAc/CyH 1+3 to
obtain 2',3',4'-Trimethoxy-biphenyl-3-carboxylic acid (34) as a
white solid (392 mg, 72%). .sup.1H NMR (400 MHz, CD.sub.3OD: 3.68
(s, 3 H); 3.93 (br.s, 6 H); 6.92 (d, 1 H, J=8.6 Hz); 7.11 (d, 1
H,J=8.6 Hz); 7.54 (t, 1 H,J=7.7 Hz); 7.75 (d, 1 H, J=7.6 Hz); 8.01
(d, 1 H, J=7.8 Hz); 8.18 (br.s 1 H).
[0115] Step 3: (The following reaction is done in an anhydrous
N.sub.2 atmosphere.) Dissolve
2',3',4'-Trimethoxy-biphenyl-3-carboxylic acid (34) (107 mg, 0.37
mmol) in anhydrous DCM (3.0 mL) and add anhydrous DMF (3 drops,
cat. amount). Then add slowly oxalyl chloride (424, 0.48 mmol) by
keeping temperature at ca. 15.degree. C. with a water bath and stir
the turbid mixture for additional 2 h at rt. Transfer the formed
crude solution of 2',3',4'-Trimethoxy-biphenyl-3-carbonyl chloride
(35) to an ice cooled solution of
[5-(2-Amino-phenyl)-thiophen-2-yl]-acetic acid methyl ester (36)
(70 mg, 0.28 mmol) in anhydrous DCM (4.5 mL) and anhydrous pyridine
(0.75 mL). Stir the reaction mixture for 3 h at it Pour the
reaction mixture into ice cooled 1M aqu. HCl, extract with DCM
(3.times.), wash the combined organic layer with brine and dry with
Na.sub.2SO.sub.4. Purify the crude product by preparative radial
chromatography (silica gel, EtOAc/CyH 1+3, later 1+2) to obtain
(5-{2-(2-[(2',3',4'-Trimethoxy-biphenyl-3-carbonyl)-amino]-phenyl}-thioph-
en-2-yl)-acetic acid methyl ester (37) as a brownish sticky solid
(96 mg, 65%). NMR (400 MHz, CDCl.sub.3): 3.64 (s, 3 H);
[0116] 3.71 (s, 3 H); 3.84 (s, 2 H); 3.90 (s, 3 H); 3.92 (s, 3 H);
6.75 (d, 1 H, J=8.8 Hz); 6.97 (d, 1 H, J=3.5 Hz); 7.01 (d, 1 H,
J=8.8 Hz); 7.03 (d, 1 H, J=3.5 Hz); 7.16 (br.t, 1 H, J=7.6 Hz);
7.36-7.43 (m, 2 H); 7.46 (t, 1 H, J=7.711z); 7.67 (.PSI.dd, 2 H,
J.sub.1=7.6 Hz, J.sub.2 =1.5 Hz); 7.91 (br.s 1 H); 8.41 (br.s 1 H);
8.50 (d, 1 H, J=8.6 Hz).
EXAMPLE 4
5-{2-Amino-4-[2-(3,4,5-trimethoxy-phenyl)-acetylamino]-phenyl}-2-methyl-fu-
ran-3-carboxylic acid (42)
##STR00046##
[0118] Step 1: (The following reaction is done under exclusion of
light.) Dissolve 2-Methyl-furan-3-carboxylic acid methyl ester (15)
(2.00 mL, 15.9 mmol) in chloroform (11 mL) and glacial acetic acid
(11 mL) and add NBS (3.85 g, 21.6 mmol) portionwise in between a
period of 75 min. Stir the reaction suspension for additional 16 h
at rt. Add water to the reaction mixture and extract the aqu. layer
with DCM (2 times), wash the combined organic layer with 2 M aqu.
NaOH, water and brine and dry it with Na.sub.2SO.sub.4 to obtain
5-Bromo-2-methyl-furan-3-carboxylic acid methyl ester (16) (2.80 g,
80%) as a red brown oil. No further purification. .sup.1H NMR (400
MHz, CDCl.sub.3): 2.54 (s, 3 H); 3.80 (s, 3 H); 6.53 (s, 1 H).
[0119] Step 2: (The following reaction is done in a N.sub.2
atmosphere.) Dissolve PdCl.sub.2(dppf) CH.sub.2Cl.sub.2 (245 mg,
0.30 mmol), KOAc (2.52 g, 25.7 mmol) and Bis-(pinacolato)diboron
(3.81 g, 15.00 mmol) in anhydrous DMSO (50 mL) and add
4-Bromo-3-nitro-phenylamine (38) (2.17 g, 10.00 mmol). Degas the
mixture carefully and flush with N.sub.2 again (5 times) and stir
it for 24 h at 80.degree. C. Cool the reaction mixture to rt and
partition it between water and toluene. Extract the aqu. layer with
EtOAc (3 times), wash the combined organic layer with water and
brine and dry it with Na.sub.2SO.sub.4. The obtained crude residue
is filtrated through a short pad of silica gel using EtOAc/CyH
(1+1) to obtain
3-Nitro-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenylami-
ne (39) (2.04 g, 77%) as a dark red solid. No further purification.
.sup.1H NMR (400 MHz, CDCl.sub.3): 1.37 (s, 12 H); 3.95 (br.s, 2
H); 6.87 (dd, 1 H, J.sub.1=7.8 Hz; J.sub.2=7.30 (d, 1 H, J=8.1 Hz);
7.35 (d, 1 H, J=2.3 Hz).
[0120] Step 3: (The following reaction is done in a N.sub.2
atmosphere.) Dissolve Pd(PPh.sub.3).sub.4 (59 mg, 0.05 mmol) and
5-Bromo-2-methyl-furan-3-carboxylic acid methyl ester (23) (447 mg,
2.04 mmol) in DME (3 mL) and stir for 10 min at rt. Add
3-Nitro-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenylamine
(39) (465 mg, 1.76 mmol) followed by an aqu. 1M sodium bicarbonate
solution (5.10 mL, 5.10 mmol). Degas the reaction mixture
carefully, flush with N.sub.2 (5 times) and stir for 4.5 h at
90.degree. C. (reflux). Cool reaction mixture to rt, remove organic
solvent under reduced pressure and partition the residue between
water and EtOAc. Extract the aqu. layer with EtOAc (3 times), wash
the combined organic layer with water and brine and dry it with
Na.sub.2SO.sub.4. Purify the obtained crude product by flash
chromatography (silica gel, EtOAc/CyH 1+3, later 1+2) to obtain
5-(4-Amino-2-nitro-phenyl)-2-methyl-furan-3-carboxylic acid methyl
ester (40) (167 mg, 34%) as a red solid. .sup.1H NMR (400 MHz,
CDCl.sub.3): 2.57 (s, 3 H); 3.81 (s, 3 H); 4.05 (br.s, 2 H); 6.68
(s, 1 H); 6.81 (dd, 1 H, J,=8.3 Hz, J.sub.2=2.3 Hz); 6.99 (d, 1 H,
J=2.3 Hz); 7.39 (d, 1 H, J=8.3 Hz).
[0121] Step 4: (The following reaction is done in an anhydrous
N.sub.2 atmosphere.) Suspend EDC.HCl (138 mg, 0.72 mmol) and
Et.sub.3N (101 .mu.L, 0.72) in anhydrous DCM (4.5 mL) and stir the
resulting solution for 5 min at rt. Add
2-(3,4,5-Trimethoxy-phenyl)-acetic acid (163 mg, 0.72 mmol) and
DMAP (8 mg, 0.07 mmol) and stir the resulting solution for 10 min.
Add 5-(4-Amino-2-nitro-phenyl)-2-methyl-furan-3-carboxylic acid
methyl ester (40) (100 mg, to 0.36 mmol) and stir the reaction
solution for 22 h at rt. Quench reaction solution with sat. aqu.
NH.sub.4Cl and water, separate layers and extract aqu. layer with
DCM (3 times). Wash the combined organic layer with water and brine
and dry with Na.sub.2SO.sub.4. Purify the crude product by
preparative radial chromatography (silica gel, EtOAc/CyH 1+1) to
obtain 2-Methyl-5-
{2-nitro-4-[2-(3,4,5-trimethoxy-phenyl)-acetylamino]-phenyl}-furan-3-carb-
oxylic acid methyl ester (41) (96 mg, 55%) as an yellow solid.
.sup.1H NMR (400 MHz, CDCl.sub.3): 2.55 (s, 3 H); 3.65 (s, 2 H);
3.79 (s, 3 H); 3.81 (s, 6 H); 3.82 (s, 3 H); 6.50 (s, 2 H); 6.77
(s, 1 H); 7.53 (d, 1 H, J=8.6 Hz); 7.66 (dd, 1 H, J.sub.1=8.6 Hz,
J.sub.2=2.0 Hz); 7.93 (br.s, 1 H); 7.96 (d, 1 H, J=2.0 Hz).
[0122] Step 5: Dissolve 2-Methyl-5-
{2-nitro-4-[2-(3,4,5-trimethoxy-phenyl)-acetylamino]-phenyl}-furan-3-carb-
oxylic acid methyl ester (41) (50 mg, 0.10 mmol) in THF (1.0 mL)
and MeOH (0.5 mL) at rt and add 1M aqu LiOH (525 .mu.L, 0.52 mmol).
Stir the reaction mixture for 17 h at rt. Add dropwise 1M aqu. HCl
(580 .mu.L, 0.58 mmol) and extract the mixture with EtOAc (3
times), wash the combined organic layer with brine and dry it with
Na.sub.2SO.sub.4. Purify the obtained crude product by preparative
TLC (silica gel, EtOAc/MeOH 9+1) to obtain
2-Methyl-5-(2-nitro-4-[2-(3,4,5-trimethoxy-phenyl)-acetylamino]-phenyl)-f-
uran-3-carboxylic acid (42) (35 mg, 71%) as a brown sticky solid.
.sup.1H NMR (400 MHz, CDCl.sub.3): 2.61 (s, 3 H); 3.70 (s, 2 H);
3.86 (s, 3 H); 3.87 (s, 6 H); 6.51 (s, 2 H); 6.85 (s, 1 H); 7.29
(br.s, 1 H); 7.58 (d, 1 H, J=8.6 Hz); 7.62 (dd, 1 H, J.sub.1=9.0
Hz, J.sub.2=2.2 Hz); 7.98 (d, 1 H, J=2.0 Hz).
EXAMPLE 5
{4-[2-(3,4,5-Trimethoxy-phenyl)-acetyl]-piperazia-1-yl}-acetic acid
ethyl ester (44)
##STR00047##
[0124] (The following reaction is done in an anhydrous N.sub.2
atmosphere.) Suspend EDC.HCl (188 mg, 0.98 mmol) and Et.sub.3N (137
.mu.L, 0.98 mmol) in anhydrous DCM (1.0 mL) and stir the resulting
solution for 5 min at rt. Add 2-(3,4,5-Trimethoxy-phenyl)-acetic
acid (163 mg, 0.72 mmol) and DMAP (8 mg, 0.07 mmol) and stir the
resulting solution for 10 min. Add
1-(Ethoxycarbonylmethyl)piperazine (43) (112 mg, 0.65 mmol) and
stir the reaction solution overnight at rt. Quench reaction
solution with sat. aqu. NH.sub.4Cl and water, separate layers and
extract aqu. layer with DCM (3 times). Wash the combined organic
layer with water and brine and dry with Na.sub.2SO.sub.4. Purify
the crude product by preparative radial chromatography (silica gel,
EtOAc/MeOH 10+1) to obtain
{4-[2-(3,4,5-Trimethoxy-phenyl)-acetyl]-piperazin-1-yl}-acetic acid
ethyl ester (44) (99 mg, 40%) as a colorless oil. .sup.1H NMR (400
MHz, CDCl.sub.3): 1.25 (t, 3 H, J=7.1 Hz); 2.48 (br.m, 2 H); 2.58
(br.m, 2 H); 3.21 (br.s, 2 H); 3.53 (br.m, 2 H); 3.65 (s, 2 H);
3.71 (br.m, 2 H); 3.81 (s, 3 H); 3.82 (s, 6 H); 4.16 (q, 2 H, J=7.1
Hz); 6.42 (s, 2 H).
EXAMPLE 6
{4-[3-(3,4,5-Trimethoxy-phenyl)-propionyl]-piperazin-1-yl}-acetic
acid (46)
##STR00048##
[0126] Step 1: (The following reaction is done in an anhydrous
N.sub.2 atmosphere.) Suspend EDC.HCl (376 mg, 1.96 mmol) and
Et.sub.3N (275 .mu.L, 1.96 mmol) in anhydrous DCM (2.0 mL) and stir
the resulting solution for 5 min at rt. Add
3-(3,4,5-Trimethoxy-phenyl)-propionic acid (346 mg, 1.44 mmol) and
DMAP (17 mg, 0.14 mmol) and stir the resulting solution for 15 min.
Add 1-(Ethoxycarbonylmethyl)piperazine (43) (224 mg, 1.30 mmol) and
stir the reaction solution overnight at rt. Quench reaction
solution with water, separate layers and extract aqu. layer with
EtOAc (3 times). Filtrate the combined organic layer through a
short pad of silica gel and remove solvent. Purify the crude
product by preparative radial chromatography (silica gel,
EtOAc/MeOH 9+1) to obtain
{4-[3-(3,4,5-Trimethoxy-phenyl)-propionyl]-piperazin-1-yl}-acetic
acid ethyl ester (45) (426 mg, 83%) as a colorless oil. .sup.1H NMR
(400 MHz, CDCl.sub.3): 1.26 (t, 3 H, J=7.1 Hz); 2.45-2.70 (br.m, 6
H); 2.89 (t, 2 H, J=7.7 Hz); 3.26 (br.s, 2 H); 3.43-3.56 (br.m, 2
H); 3.61-3.76 (br.m, 2 H); 3.80 (s, 3 H); 3.83 (s, 6 H); 4.18 (q, 2
H, J=7.1 Hz); 6.41 (s, 2 H).
[0127] Step 2: Dissolve
{4-[3-(3,4,5-Trimethoxy-phenyl)-propionyl]-piperazin-1-yl}-acetic
acid ethyl ester (45) (100 mg, 0.25 mmol) in MeOH (2.0 mL) at rt
and add 2M aqu NaOH (260 .mu.L, 0.52 mmol). Stir the reaction
mixture for 1 h under reflux. Add dropwise 1M aqu. HCl (550 .mu.L,
0.55 mmol), extract the mixture with EtOAc (3 times) and remove
solvent obtain
{4-[3-(3,4,5-Trimethoxy-phenyl)-propionyl]-piperazin-1-yl}-acetic
acid (46) (88 mg, 95%) as a brown sticky solid. No further
purification. .sup.1H NMR (400 MHz, CDCl.sub.3/CD.sub.3OD 9+1):
2.54 (br.t, 2 H); 2.78 (t, 2 H, J=7.5 Hz); 2.83-3.10 (br.m, 2 H);
3.24 (s, 2 H); 3.43-3.62 (br.m, 2 H); 3.68 (s, 3 H); 3.73 (s, 6 H);
3.74-3.85 (br.m, 4 H); 6.34 (s, 2 H).
EXAMPLE 7
{2'-[3-(3,4,5-Trimethoxy-phenyl)-propionylamino]-biphenyl-3-yl}-acetic
acid methyl ester (50)
##STR00049##
[0129] Step 1: (The following reaction is done in an oxygenfree
N.sub.2 atmosphere.) Add ethanol (0.8 mL),
Tetrakis-(triphenylphosphine)-palladium(0) (30 mg, 2.2 mol %) and
Na.sub.2CO.sub.3 decahydrate (944 mg, 3.30 mmol; presolved in 1.2
mL H.sub.2O) subsequently to dissolved 2-Amino-benzeneboronic acid
(48) (201 mg, 1.30 mmol) in toluene (6.0 mL). Degas the reaction
mixture for 5 times and flood with N.sub.2 again. Add
(3-Bromo-phenyl)-acetic acid methyl ester (47) (270 mg, 1.18 mmol)
in toluene (6.0 mL), degas again (5 times) and stir the reaction
solution overnight at 100.degree. C. Partition the reaction
solution between EtOAc and brine (1+1) and extract the separated
aqueous layer 3 times with EtOAc. Wash combined organic layer with
brine and dry with Na.sub.2SO.sub.4. Remove solvent under reduced
pressure and purify the crude product by preparative radial
chromatography (silica gel 60PF, CyH/EtOAc 3+1) to obtain
(2'-Amino-biphenyl-3-yl)-acetic acid methyl ester (49) as an orange
oil (304 mg, 81%). .sup.1H NMR (400 MHz, CDCl.sub.3): 3.66 (s, 2
H); 3.69 (s, 3 H); 3.62-3.86 (br.s, 2 H); 6.75 (d, 1 H, J=8.1 Hz);
6.80 (t, 1 H, J=7.3 Hz); 7.11 (d, 1 H, J=7.3 Hz); 7.15 (d, 1 H,
J=8.1 Hz); 7.22-7.26 (br.m, 1 H); 7.32-7.42 (m, 3 H).
[0130] Step 2: (The following reaction is done in an anhydrous
N.sub.2 atmosphere.) Suspend EDC.HCl (61 mg, 0.32 mmol) and
Et.sub.3N (44 .mu.L, 0.32 mmol) in anhydrous DCM (1.0 mL) and stir
the resulting solution for 5 min at rt. Add
3-(3,4,5-Trimethoxy-phenyl)-propionic acid (55 mg, 0.23 mmol) and
DMAP (2 mg, 0.02 mmol) and stir the resulting solution for 15 min.
Add (2'-Amino-biphenyl-3-yl)-acetic acid methyl ester (49) (50 mg,
0.21 mmol) and stir the reaction solution overnight at rt. Quench
reaction solution with water, separate layers and extract aqu.
layer with DCM (3 times). Wash combined organic layer with brine
and dry with Na.sub.2SO.sub.4. Purify the crude product by
preparative radial chromatography (silica gel, EtOAc/CyH 1+1) to
obtain
{2'-[3-(3,4,5-Trimethoxy-phenyl)-propionylamino]-biphenyl-3-yl}-acetic
acid methyl ester (50) (46 mg, 48%) as a yellow oil. .sup.1H NMR
(400 MHz, CDCl.sub.3): 2.50 (t, 2 H, J=7.6 Hz); 2.90 (t, 2 H, J=7.7
Hz); 3.64 (s, 2 H); 3.65 (s, 3 H); 3.77 (s, 6 H); 3.78 (s, 3 H);
6.38 (s, 2 H); 7.09-7.18 (m, 3 H); 7.19-7.28 (m, 3 H); 7.34 (d, 1
H, J=8.1 Hz); 7.38 (d, 1 H, J=7.8 Hz); 8.31 (br.d, 1 H, J=7.8
Hz).
EXAMPLE 8
4-[3-(3,4,5-Trimethoxy-phenyl)-propionylamino]-benzoic acid methyl
ester (52)
##STR00050##
[0132] (The following reaction is done in an anhydrous N.sub.2
atmosphere.) Suspend EDC.HCl (80 mg, 0.41 mmol) and Et.sub.3N (58
.mu.L, 0.41 mmol) in anhydrous DCM (2.0 ML) and stir the resulting
solution for 5 min at rt. Add 3-(3,4,5-Trimethoxy-phenyl)-propionic
acid (70 mg, 0.29 mmol) and DMAP (5 mg, 0.04 mmol) and stir the
resulting solution for 10 min. Add 4-Amino-benzoic acid methyl
ester (51) (42 mg, 0.27 mmol) and stir the reaction solution 2 d at
rt. Quench reaction solution with water, separate layers and
extract aqu. layer with DCM (3 times). Wash combined organic layer
with brine, dry with Na.sub.2SO.sub.4 and filtrate it through a
short pad of silica gel using EtOAc to obtain
4-[3-(3,4,5-Trimethoxy-phenyl)-propionylamino]-benzoic acid methyl
ester (52) (91 mg, 88%) as a white solid. No further purification.
.sup.1H NMR (400 MHz, CDCl.sub.3): 2.60 (t, 2 H, J=7.6 Hz); 2.91
(t, 2 H, J=7.6 Hz); 3.70 (s, 6 H); 3.76 (s, 3 H); 3.83 (s, 3 H);
6.35 (s, 2 H); 7.55 (d, 2 H, J=8.3 Hz); 7.91 (d, 2 H, J=8.6 Hz);
8.09 (s, 1 H).
EXAMPLE 9
(5-{2-[(3',4',5'-Trimethoxy-biphenyl-2-carbonyl)-amino]-phenyl}-thiophen-2-
-yl)-acetic acid (59)
##STR00051## ##STR00052##
[0134] Step 1: (The following reaction is done in an N.sub.2
atmosphere.) To a solution of Methyl-2-bromobenzoate (53) (922 mg,
4.29 mmol) in toluene (11 mL) is added Pd(PPh.sub.3).sub.4 (297 mg,
0.26 mmol) and Na.sub.2CO.sub.310H.sub.2O (3.43 g, 12.00 mmol) in
water (3.8 mL). Degas the resulting mixture is carefully (5 times
alternating vacuum and flushing with N.sub.2). Add a solution of
3,4,5-Trimethoxyphenylboronic acid (54) (1.00 g, 4.72 mmol) in
toluene (10 mL) by syringe, degas the resulting mixture again
carefully and stir the resulting mixture overnight at 100.degree.
C. Partition the mixture between brine/EtOAc (1+1), separate
layers, extract the aqu. layer with EtOAc (3.times.), wash the
combined organic layer with brine and dry with Na.sub.2SO.sub.4.
Purify the crude product by flash chromatography (silica gel,
EtOAc/CyH 1+7, later 1+5) to obtain
3',4',5'-Trimethoxy-biphenyl-2-carboxylic acid methyl ester (55) as
a yellow solid (1.30 g, 99%). .sup.1H NMR (400 MHz, CDCl.sub.3):
3.65 (s, 3 H); 3.84 (s, 6 H); 3.87 (s, 3 H); 6.52 (s, 2 H);
7.35-7.42 (m, 2 H); 7.50 (t, 1 H, J=8.0 Hz); 7.73 (d, 1 H, J=8.0
Hz).
[0135] Step 2: Dissolve 3',4',5'-Trimethoxy-biphenyl-2-carboxylic
acid methyl ester (55) (626 mg, 2.08 mmol) in MeOH (14 mL) at rt
and add 1M aqu LiOH (4.2 mL, 4.20 mmol). Stir reaction mixture for
8 h under reflux. Remove solvent and partition the residue between
1M aqu. HCl and EtOAc, separate layers, extract the aqu. layer with
EtOAc (3.times.), wash the combined organic layer with brine and
dry with Na.sub.2SO.sub.4. Remove solvent and recrystallize residue
from EtOAc/CyH 1+2 to obtain
3',4',5'-Trimethoxy-biphenyl-2-carboxylic acid (56) as a white
solid (423 mg, 79%). .sup.1H NMR (400 MHz, CD.sub.3OD: 3.84 (s, 3
H); 3.89 (s, 6 H); 6.68 (s, 2 H); 7.42 - 7.49 (m, 2 H); 7.57 (t, 1
H, J=7.5 Hz); 7.76 (d, 1 H, J=8.014z).
[0136] Step 3: (The following reaction is done in an anhydrous
N.sub.2 atmosphere.) Dissolve
3',4',5'-Trimethoxy-biphenyl-2-carboxylic acid (56) (54 mg, 0.18
mmol) in anhydrous DCM (1.3 mL) and add anhydrous DMF (1 drop, cat.
amount). Then add slowly oxalyl chloride (21 .mu.L, 0.24 mmol) by
keeping temperature at ca. 20.degree. C. with a water bath and stir
the turbid mixture for additional 2 h at it Remove solvent and dry
in vacuum to obtain crude 3',4',5'-Trimethoxy-biphenyl-2-carbonyl
chloride (57) as a yellow solid. No further purification.
[0137] Step 4: Add a solution of
3',4',5'-Trimethoxy-biphenyl-2-carbonyl chloride (57) (0.18 mmol)
in DCM (1.0 mL) to an ice cooled solution of
[5-(2-Amino-phenyl)-thiophen-2-yl]-acetic acid methyl ester (36)
(46 mg, 0.18 mmol) in anhydrous DCM (2.0 mL) and anhydrous pyridine
(0.5 mL). Stir the reaction mixture for 1 h at 0.degree. C. and
additional 20 h at rt. Pour the reaction mixture into ice cooled 1M
aqu. HCl, extract with EtOAc (2.times.) and DCM (2.times.), wash
the combined organic layer with brine and dry with
Na.sub.2SO.sub.4. Purify the crude product by preparative radial
chromatography (silica gel, EtOAc/CyH 1+2) to obtain
(5-{2-[(3',4',5'-Trimethoxy-biphenyl-2-carbonyl)-amino]-phenyl}-thiophen--
2-yl)-acetic acid methyl ester (58) as a light brown solid (58 mg,
59%). .sup.1H NMR (400 MHz, CDCl.sub.3): 3.70 (s, 3 H); 3.76 (s, 6
H); 3.78 (s, 2 H); 3.80 (s, 3 H); 6.29 (d, 1 H, J=3.4 Hz); 6.60 (s,
2 H); 6.75 (d, 1 H, J=3.4 Hz); 7.07 (t, 1 H, J=7.6 Hz); 7,23 (d, 1
H, J=7.6 Hz); 7.31 (t, 1 H, J=8.0 Hz); 7.37 -7.43 (m, 2 H); 7.48
(t, 1 H, J=7.6 Hz); 7.52 (s, 1 H); 7.69 (d, 1 H, J=8.0 Hz); 8.45
(d, 1 H, J=8.0 Hz).
[0138] Step 5: Dissolve (5-{2-[(3',4',540
-Trimethoxy-biphenyl-2-carbonyl)-amino]-phenyl}-thiophen-2-yl)-acetic
acid methyl ester (58) (56 mg, 0.11 mmol) in MeCN (3.8 mL) at rt
and add 1M aqu LiOH (760 .mu.L, 0.76 mmol). Stir reaction mixture
18 h at rt. Quench reaction mixture (cooling bath) with 2 M aqu.
HCl. Extract the mixture with EtOAc (3.times.), wash the combined
organic layer with brine and dry with Na.sub.2SO.sub.4 to obtain
(5-{2-[(3',4',5'-Trimethoxy-biphenyl-2-carbonyl)-amino]-phenyl}-thiophen--
2-yl)-acetic acid (59) (55 mg, 99%) as a brown solid. .sup.1H NMR
(400 MHz, CDCl.sub.3): 3.76 (s, 6 H), 3.80 (s, 3 H); 3.83 (s, 2 H);
6.32 (d, 1 H, J=3.5 Hz); 6.60 (s, 2 H); 6.78 (d, 1 H, J=3.5 Hz);
7.07 (t, 1 H, J=7.6 Hz); 7.23 (d, 1 H, J=7,6 Hz); 7.32 (t, 1 H,
J=7.6 Hz); 7.36-7.54 (m, 3 H); 7.69 (d, 1 H, J=8.0 Hz); 8.43 (d, 1
H, J=8.0 Hz).
EXAMPLE 10
2',3',4'-Trimethoxy-biphenyl-3-carboxylic acid
{2-[5-(1H-tetrazol-5-ylmethyl)-thiophen-2-yl]-phenyl}-amide
(87)
##STR00053##
[0140] Step 1: (The following reaction is done in an anhydrous
N.sub.2 atmosphere.) Dissolve nitrile (82) (500 mg, 4.06 mmol) in
anhydrous DMF (2.7 mL), cool to 0.degree. C., add
N-bromosuccinimide (795 mg, 7.79 mmol) in portions over a period of
20 min and stir the mixture for 22 h at it Partition the reaction
solution between dichloromethane and water (1+1) and extract the
separated aqueous layer 2 times with dichloromethane. Wash combined
organic layer with water and brine and dry with Na.sub.2SO.sub.4.
Remove solvent under reduced pressure and purify the crude product
by preparative radial chromatography (silica gel 60PF, CyH/EtOAc
10+1] to obtain (5-bromo-thiophen-2-yl)-acetonitrile (83) as a
light yellow liquid (745 mg, 91%). [M. A. Ismail, R. Brun, J. D.
Easterbrook, F. A. Tanious, W. D. Wilson, D. W. Boykin, J. Med
Chem. 2003; 46 (22); 4761-4769]. .sup.1H NMR (400 MHz, CDCl.sub.3):
3.81 (d, 2 H, J=1.0 Hz); 6,81 (d, 1 H, J.sub.1=3.8 Hz, J.sub.2=1.0
Hz); 6.92 (d, 1 H, J=3.8 Hz).
[0141] Step 2: (The following reaction is done in an oxygenfree
N.sub.2 atmosphere.) Dissolve
tetrakis-(triphenylphosphine)-palladium(0) (29 mg, 2.5 mol %) and
nitrile (83) (101 mg, 0.50 mmol) in DME (3.7 mL). The reaction
mixture is degassed 5 times and flooded with N.sub.2 again. Add
2-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)phenylamine (84)
(120 mg, 0.55 mmol), rinse with DME (0.5 mL), add aqu. 1M
NaHCO.sub.3, degas again (5 times) and stir the reaction solution 2
h at 90.degree. C. (reflux). After cooling to rt partition the
reaction mixture between EtOAc and brine (1+1) and extract the
separated aqueous layer 3 times with EtOAc. Wash combined organic
layer with brine and dry with Na.sub.2SO.sub.4. Remove solvent
under reduced pressure and purify the crude product by preparative
radial chromatography (silica gel 60PF, CyH/EtOAc 2+1) to obtain
[5-(2-amino-phenyl)-thiophen-2-yl]-acetonitrile (85) as a brownish
oil (84 mg, 78%). .sup.1H NMR (400 MHz, CDCl.sub.3): 3.89 (br.s, 2
H); 5.20-6.50 (br.s, 2 H); 6.92 (t, 1 H; J=7.6 Hz); 6.95-7.00 (m, 1
H); 6.99 (d, 1 H; J=3.8 Hz); 7.11 (d, 1 H; J=3.8 Hz); 7.21 (dd, 1
H; J.sub.1=7.6 Hz, J.sub.2=1.3 Hz); 7.27 (dd, 1 H; J.sub.1=7.6 Hz,
J.sub.2=1.3 Hz).
[0142] Step 3: (The following reaction is done in an anhydrous
N.sub.2 atmosphere.) Add a solution of
2',3',4'-trimethoxy-biphenyl-3-carbonyl chloride (35) (117 mg, 0.38
mmol) in dichloromethane (1.3 mL) to an ice cooled solution of the
aniline (85) (82 mg, 0.38mmol) in anhydrous dichloromethane (2.6
mL) and anhydrous pyridine (0.65 mL). Stir the reaction mixture for
1 h at 0.degree. C. and additional 21 h at rt. Pour the reaction
mixture into ice cooled 1M aqu. HCl (20 mL), extract with
dichloromethane (2.times.) and EtOAc (1.times.), wash the combined
organic layer with brine and dry with Na.sub.2SO.sub.4. Purify the
crude product by preparative radial chromatography (silica gel
60PF, EtOAc/CyH 1+3 (incl. 2% MeOH), later 1+3) to obtain anilide
(86) as an orange solid (132 mg, 71%). .sup.1H NMR (400 MHz,
C.sub.6D.sub.6) 2.88 (d, 2 H, J=1.0 Hz); 3.53 (s, 3 H); 3.55 (s, 3
H), 3.94 (s, 3 H), 6.63 (d, 1 H, J=3.5 Hz); 6.64 (d, 1 H, J=8.6
Hz); 6.66 (d, 1 H, J=3.5 Hz); 6.99 (td, 1 H; J.sub.1=7.6 Hz,
J.sub.2=1.0 Hz); 7.01 (d, 1 H, .sub.J=8.6 Hz), 7.25-7.34 (m, 3 H);
7.74 (dt, 1 H; J.sub.1=7.8 Hz, J.sub.2=1.5 Hz); 7.97 (dt, 1 H,
J.sub.1=7.8 Hz, J.sub.2=1.5 Hz), 8.18 (t, 1 H, J=1.5 Hz), 8.34 (br.
s, 1 H); 9.14 (d, 1 H, J=7.8 Hz).
[0143] Step 4: (The following reaction is done in an anhydrous
N.sub.2 atmosphere.) Dissolve anilide (86) (60 mg, 0.12 mmol) in
anhydrous DMF (2.0 mL), add sodium azide (18 mg, 0.14 mmol) and
ammonium chloride (9 mg, 0.85 mmol) and stir the reaction solution
for 2 d at 90.degree. C. Add again sodium azide (18 mg, 0.14 mmol)
and ammonium chloride (9 mg, 0.85 mmol) and stir for additional 3d
at 90.degree. C. Cool mixture to rt and adjust pH=1 by addition of
1M HCl. Extract aqueous layer with dichloromethane (3.times.). Wash
the combined organic layer with brine, dry with Na.sub.2SO.sub.4
and remove solvent under reduced pressure. Purify the crude product
by preparative radial chromatography (silica gel, EtOAc/CyH 1+2,
later EtOAc/MeOH 9+1) to obtain
2',3',4'-trimethoxy-biphenyl-3-carboxylic acid
{2-[5-(1H-tetrazol-5-ylmethyl)-thiophen-2-yl]-phenyl}-amide (87) as
a yellow solid (50 mg, 76%) [F. Osterod, L. Peters, A. Kraft, T.
Sano, J. J. Morisson, N. Feeder, A. B. Holmes, J. Mater. Chem.
2001, 11, 1625-1633 and refer. therein. .sup.1H-NMR. (400 MHz,
(CD.sub.3).sub.2SO): 3.61 (s, 3 H); 3.80 (s, 3 H); 3.84 (s, 3 H);
4.45 (s, 2 H); 6.93 (d, 1 H; J=8.6 Hz); 6.93 (d, 1 H; J=3.5 Hz);
7.11 (d, 1 H, J=8.6 Hz); 7.26 (d, 1 H, J=3.5 Hz); 7.33 (td, 1 H,
J.sub.1=7.3 Hz, J.sub.2=1.8 Hz); 7.35 (td, 1 H, J.sub.1=7.3 Hz,
J.sub.2=1.8 Hz); 7.43-7.47 (m, 1 H); 7.53 (t, 1 H, J=7.7 Hz); 7.61
(dd, 1 H, J.sub.1=7.0 Hz, J.sub.2=2.0 Hz); 7.66 (d, 1 H, J=7.6 Hz);
7.85 (d, 1 H, J=7.6 Hz); 7.99 (s, 1 H); 10.07 (s, 1 H).
EXAMPLE 11
2',3',4'-Trimethoxy-biphenyl-3-carboxylic acid
(2-bromo-5-nitro-phenyl)-amide (89)
##STR00054##
[0144] (The following reaction is done in an anhydrous N.sub.2
atmosphere.) Add a solution of
2',3',4'-trimethoxy-biphenyl-3-carbonyl chloride (35) (345 mg, 1.13
mmol) in dichloromethane (5 mL) to an ice cooled solution of
2-bromo-5-nitro-phenylamine (88) (232 mg, 1.07 mmol) in anhydrous
dichloromethane (12 mL) and anhydrous pyridine (2.9 mL). Stir the
reaction mixture for 15 min at 0.degree. C. and additional 17 h at
rt. Pour the reaction mixture into ice cooled 1M aqu. HCl (to get
pH ca. 3), extract with EtOAc (3.times.), wash the combined organic
layer with brine and dry it with Na.sub.2SO.sub.4 to afford crude
2',3',4'-trimethoxy-biphenyl-3-carboxylic acid
(2-bromo-5-nitro-phenyl)-amide (89) as a beige solid (542 mg,
quant.). NMR (400 MHz, CDCl.sub.3): 3.72 (s, 3 H); 3.91 (s, 3 H);
3.93 (s, 3 H); 6.77 (d, 1 H, J=8.6 Hz); 7.08 (d, 1 H, J=8.6 Hz);
7.56 (t, 1 H, J=8.8 Hz); 7.75 (d, 2 H, J=8.8 Hz); 7.86 (d, 1 H,
J=8.8 Hz), 7.87 (d, 1 H, J=8.8 Hz); 8.09 (t, 1 H, J=1.7 Hz); 8.61
(br. s, 1 H); 9.59 (d, 1 H, J=2.5 Hz).
EXAMPLE 12
N-(3-Nitro-phenyl)-3-(3,4,5-trimethoxy-phenyl)-propionamide
(91)
##STR00055##
[0146] (The following reaction is done in an anhydrous N.sub.2
atmosphere.) Suspend EDC hydrochloride (402 mg, 2.10 mmol) and
Et.sub.3N (293 .mu.L, 2.10 mmol) in anhydrous dichloromethane (17.0
mL) and stir the resulting solution for 5 min at rt. Add
3-(3,4,5-trimethoxy-phenyl)-propionic acid (481 mg, 2.00 mmol) and
DMAP (24 mg, 0.20 mmol) and stir the resulting solution for 5 min.
Add 3-nitro-phenylamine (90) (414 mg, 3.00 mmol) and stir the
reaction solution 24 h at rt. Quench reaction solution with sat.
aqu. NH.sub.2Cl and water, separate layers and extract aqu. layer
with EtOAc (3 times). Wash the combined organic layer with water
and brine and dry with Na.sub.2SO.sub.4. Purify the crude product
by preparative radial chromatography (silica gel 60PF, EtOAc/CyH
1+1) to obtain
N-(3-nitro-phenyl)-3-(3,4,5-trimethoxy-phenyl)-propionamide (91)
(508 mg, 70%) as a yellowish solid. .sup.1H NMR (400 MHz,
CDCl.sub.3): 2.65 (t, 2 H, J=7.3 Hz); 2.98 (t, 2 H, J=7.3 Hz); 3.79
(s, 6 H); 3.81 (s, 3 H), 6.42 (s, 2 H); 7.41 (s, 1 H); 7.45 (t, 1
H, J=8.0 Hz); 7.84 (d, 1 H, J=8.0 Hz); 7.92 (d, 1 H, J=8.01 Hz);
8.31 (s, 1 H).
[0147] The compounds referred to in the following SCHEME 16 are
those compounds referred to as the particularly preferred compounds
herein.
##STR00056## ##STR00057## ##STR00058## ##STR00059##
##STR00060##
Sialyl Lewis.sup.X Tyrosine Sulfate Assay (sLe.sup.x TSA):
[0148] Compounds of the present invention are assayed on a
molecular level for their ability to inhibit the binding of P-, L-,
or E-selectin chimeric molecules to sLe.sup.x and tyrosinesulfate
residues linked to a polymeric matrix as a PSGL-1 substitute.
IC.sub.50-values are determined.
[0149] Microtiter plates are coated overnight in carbonate buffer
pH9.6 with goat anti human Fc mAB (10 .mu.g/ml). After washing in
assay buffer (25 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic
acid (HEPES), 150 mM NaCl, 1 mM CaCl.sub.2 pH7.4) and blocking (3%
bovine serum albumin (BSA) in assay buffer) plates are incubated
for 2 h at 37.degree. C. with human P-Selectin-IgG-chimera (0.61 nM
respectively 150 ng/mL) or human L-Selectin-IgG-chimera (0.61 nM
respectively 89 ng/mL) or human E-Selectin-IgG-chimera (0.61 nM
respectively 131 ng/mL). 5 .mu.l of sLe.sup.x-tyrosine sulfate
polyacrylamide (1 mg/ml) carrying 15% sLe.sup.x, 10%
Tyrosine-sulfate and 5% biotin is complexed with 20 .mu.l
Streptavidin-Peroxidase solution (1 mg/ml) and 25 .mu.l assay
buffer without CaCl.sub.2. For use in the assay, the ligand complex
is diluted 1:10000 in assay buffer and further diluted 1:1 with
varying amounts of compounds in assay buffer incl. 2% DMSO. This
mixture is added to the wells precoated with E- or P-selectin.
After incubation for 2 h at 37.degree. C., wells are washed for six
times with in assay buffer incl. 0.005% Polyoxyethylenesorbitan
monolaurate (TWEEN 20), developed for 10-15 min with 20 .mu.l
3,3',5,5'-tetramethylbenzidine (TMB)/H.sub.2O.sub.2 substrate
solution and stopped with 20 .mu.l 1M H.sub.2SO.sub.4. Bound
sLe.sup.x-Tyrosine sulfate ligand complex is determined by
measuring optical density at 450 nm vs. 620nm in a Fusion alpha-FP
reader (sold from Packard Bioscience, Dreieich, Germany).
Results from sLe.sup.xTSA: IC.sub.50 Data for E-/P-/L-Selectin
TABLE-US-00001 IC.sub.50 E-Selectin IC.sub.50 P-Selectin IC.sub.50
L-Selectin Compound [.mu.M] [.mu.M] [.mu.M] Bimosiamose >500
95.0 >500 60 18.2 15.0 12.8 61 >500 186.1 385.3 62 74.7 46.4
45.3 63 >500 28.5 76.1 64 >500 107.1 382.9
Results from sLe.sup.xTSA: IC.sub.50 Data for E-/L-Selectin
TABLE-US-00002 IC.sub.50 E-Selectin IC.sub.50 P-Selectin IC.sub.50
L-Selectin Compound [.mu.M] [.mu.M] [.mu.M] 87 -- 32.6 59.5
Flow Chamber Assay/Cell Adhesion and Rolling under Flow
Conditions
[0150] To assess the capability of compounds to inhibit cell
binding under dynamic conditions resembling the flow in a blood
vessel, flow chamber assays addressing/ testing binding of HL-60
cells/various cell lines to P-selectin, L-selectin and E-selectin
chimeric molecules are performed.
[0151] Cell attachment under flow conditions are determined using a
parallel flow chamber system. A 35mm polystyrene culture dish is
coated for 1 hour at room temperature with coating buffer (50mM
tris-(hydroxymethyl) aminomethane buffer (Tris), 150 mM NaCl, 2 mM
CaCl.sub.2; pH 7.4) containing human E- or P-selectin-IgG chimera
at concentrations of 2,5 .mu.g/ml or 10 .mu.g/ml, respectively.
After removal of the coating solution non specific binding sites
are blocked for an additional hour with 1% BSA in coating buffer at
room temperature. After washing with assay buffer ("Roswell Park
Memorial Institute 1640" (RPMI 1640)+10 mM HEPES) the dish is
fitted into a parallel plate laminar flow chamber (sold from
Glycotech, Rockville, Md.) and mounted on an inverted
phase-contrast microscope (sold from Olympus, Hamburg, Germany)
equipped with a CCD camera (JVC) that is connected to a PC.
Employing a peristaltic pump (sold from Ismatec, Wertheim-Mondfeld,
Germany) the re-circulating system is equilibrated with assay
buffer containing 125 .mu.M compound or vehicle control (DMSO).
Cells (1 million/ml) are added to the chamber and allowed to
distribute for 2 minutes at a high flow rate. The flow rate is then
decreased resulting in a calculated flow shear of 1 dyne/cm.sup.2.
Video sequences of 10 low power fields are digitally recorded after
5 minutes continuous flow. The percentage of modulation is
calculated from the mean number of cells per field that attached to
the coated dish surface in the presence versus absence of compound
of at independent experiments.
Data from Flow Chamber Assay for E- and P-Selectin
[0152] Values are given as normalized ratios of %-inhibition of
compound x divided by %-inhibition of bimosiamose.
TABLE-US-00003 E-Selectin P-Selectin Compound [Ratio] [Ratio] 63
1.46 1.06 64 1.27 1.01 87 1.23 2.62
* * * * *